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PAPERS
Salmonella colonisation of laying hens
following vaccination with killed and
live attenuated commercial Salmonella
vaccines
R. J. Atterbury, J. J. Carrique-Mas, R. H. Davies, V. M. Allen

The aim of this study was to determine the efficacy of
a killed Salmonella vaccine and three live vaccines in
preventing caecal colonisation of Hy-line Brown pullets
by Salmonella Enteritidis PT 4. The lowest number of
Salmonella-positive birds following the largest challenge
(108 cfu) was recorded for live vaccine 1. However,
birds treated with the killed vaccine had a significantly
lower number of salmonellae in their caeca compared
with both the control group and the other vaccine
groups (P<0·05).
ACUTE bacterial enteritis caused by Salmonella species is a major public
health burden internationally. In 2006, there were 160,649 confirmed
cases of human salmonellosis in the EU (European Food Safety Authority
[EFSA] 2007a) and 12,506 in England and Wales, of which 54·6 per cent
were due to Salmonella Enteritidis (Health Protection Agency [HPA]
2009a). Contaminated poultry products (eggs and meat) are widely
accepted as a primary source of human S Enteritidis infection (Rabsch and
others 2001, de Jong and Ekdahl 2006), and most cases are attributed to
the consumption of contaminated eggs or egg products (Coyle and others
1988, Gillespie and others 2005). Recent large outbreaks of S Enteritidis-
associated disease in human beings in the UK have been linked with low-
cost eggs imported from continental Europe (HPA 2009b). A beneficial
effect of vaccination against Salmonella has been shown in both experi-
mental work (Timms and others 1990, Barbour and others 1993, Gast and
others 1993, Nakamura and others 1994, Curtiss and Hassan 1996, Holt
and others 1996, Linde and others 1996, Miyamoto and others 1999) and
Veterinary Record (2009) 165, 493-496
R. J. Atterbury, BSc, PhD, Dr Atterbury’s present address is
V. M. Allen, PhD, FIBMS, School of Veterinary Medicine and
Department of Clinical Veterinary Science, University of Nottingham,
Science, University of Bristol, Sutton Bonington, Leicestershire,
Langford, Bristol BS40 5LX LE12 5RD
J. J. Carrique-Mas, DVM, MSc,
PhD, MRCVS,
R. H. Davies, BVSc, PhD, MRCVS, E-mail for correspondence:
Department of Bacterial Diseases, robert.atterbury@nottingham.ac.uk
Veterinary Laboratories Agency –
Weybridge, Woodham Lane, New
Haw, Addlestone, Surrey KT15 3NB
field studies (Yamane and others 2000, Feberwee and others 2001, Davies
and Breslin 2003). In addition, an EU-wide baseline report into the prev-
alence of Salmonella in large-scale laying hen holdings reported that vac-
cination reduces the probability of S Enteritidis infection by 88 per cent
(EFSA 2007b). However, other studies have failed to demonstrate sig-
nificant reductions in colonisation with Salmonella following vaccination
(Davison and others 1999, Parker and others 2001). There are few data
on the efficacy of vaccination when layer hens are challenged with field
isolates of Salmonella under simulated commercial conditions. Two com-
mercial killed vaccines, one containing an iron-restricted S Enteritidis
PT4 bacterin vaccine, the other containing an iron-restricted bacterin
vaccine with inactivated S Enteritidis and Salmonella Typhimurium, have
previously been shown to significantly reduce the prevalence of Salmonella
in both experimental studies (Woodward and others 2002) and field stud-
ies (Feberwee and others 2001). The present study compared the efficacy
of one killed vaccine and three live vaccines currently available in the
UK in reducing caecal colonisation with a field strain of S Enteritidis in
point-of-lay pullets under simulated commercial conditions.
Materials and methods
Vaccines
The live vaccines were delivered in drinking water by reconstituting
lyophilised vials of each product in the recommended volume of water.
Water was withdrawn from the birds for a period of three hours before
they were provided with troughs containing the vaccine suspension.
The birds then consumed this suspension ad libitum over a period rec-
ommended by the manufacturer, which was not less than four hours. A
coloured dye was used in the vaccine suspension to check that all of the
birds had consumed some of the liquid.
Live vaccine 1 (LV1) consisted of 1 x 108 to 8 x 108 cfu of a double-
attenuated (adenine-histidine auxotrophic) S Enteritidis mutant (strain
441/014). The manufacturer’s recommended vaccination schedule was
followed (dose 1 at one day old, dose 2 at two weeks of age and dose 3 at
three weeks before the onset of lay). Live vaccine 2 (LV2) consisted of
1 x 108 cfu of an attenuated mutant (metabolic drift) S Enteritidis strain
Sm24/Rif12/SSq. The manufacturer’s recommended vaccination schedule
was followed (dose 1 at one day old, dose 2 at seven weeks of age and dose
3 at three weeks before the onset of lay). Live vaccine 3 (LV3) consisted
of 1 x 108 to 6 x 108 cfu of a live attenuated mutant (metabolic drift) of
S Typhimurium strain Nal2/Rif9/Rtt PT 9. The manufacturer’s recom-
mended vaccine schedule was followed (dose 1 at one day old, dose 2 at
seven weeks of age and dose 3 at three weeks before the onset of lay). LV3
was used only in conjunction with LV2 (both vaccines were manufactured
by the same company); this was done as the vaccines protected against
different serovars of Salmonella. Unpublished evidence suggested that the
combination of LV2 and LV3 was more effective against S Enteritidis
than LV2 used alone (R. H. Davies, personal communication).

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PAPERS

The killed vaccine (KV) consisted of

TABLE 1: Caecal colonisation of point-of-lay pullets following challenge with a low dose
formalin-killed cells of S Enteritidis PT4 and (1·5 x 102 cfu) or a high dose (1·5 x 108 cfu) of Salmonella Enteritidis
S Typhimurium DT104 (approximately 109
cfu of each per 0·5 ml dose). The birds were Number (%) of birds with
Number (%) of Median colonisation ≥2·0 log10 cfu Salmonella/
vaccinated twice with 0·5 ml of the KV by Vaccine used Number of birds in group S Enteritidis-positive birds (cfu/g) g caecal contents
intramuscular injection into the breast at 12
and 16 weeks of age. Low challenge dose
LV2 18 10 (55·6) – 0 (0)
LV2+LV3 18 7 (38·9) 3 1 (5·6)
Experimental design Control 1* 18 11 (61·1) 6·3 1 (5·6)
Non-vaccinated, commercial day-of-hatch LV1 18 4 (22·2) – 0 (0)
Hy-line Brown pullet chicks (n=192) were KV 18 5 (27·8) 4 1 (5·6)
separated at random into four groups of 36 Control 2† 24 8 (33·3) 2·7 1 (5·6)
High challenge dose
birds (groups A, B, C, D) and one group of
LV2 18 16 (88·9) 3·9 10 (55·5)
48 birds (group E). Group A was vaccinated LV2+LV3 18 15 (83·3) 4·9 14 (77·8)
with LV1, group B was vaccinated with LV2, Control 1* 18 17 (94·4) 4·8 16 (88·9)
group C was vaccinated with LV2 and LV3, LV1 18 13 (72·2) 3·8 9 (50·0)
group D was vaccinated with KV, and group E KV 18 14 (77·8) 3·8 1 (5·6)
remained unvaccinated for the duration of Control 2† 24 24 (100) 4·3 16 (66·7)
the trial. The groups were housed separately * Unvaccinated group challenged at the same time as the groups vaccinated with LV2 and LV3
under conditions of strict biosecurity. Swabs †
Unvaccinated group challenged at the same time as the groups vaccinated with LV1 and KV
of representative voided faeces (six swabs KV Killed vaccine, LV Live vaccine, – Unable to calculate as no samples contained directly countable numbers of Salmonella
taken from each of six pooled samples of fae-
ces) from each group of birds were taken on
a fortnightly basis before the challenge in order to detect any naturally A sterile swab was inserted into the liver of each bird and used to
occurring Salmonella infections. The swabs were used to inoculate 10 ml inoculate a plate of BG agar. The inoculum was streaked over the sur-
of Rappaport-Vassiliadis soya peptone broth (RVS, CM0866; Oxoid), face of the plate before incubation aerobically at 37°C for 24 hours.
which was incubated for 24, hours at 41·5°C and then streaked directly The swab was transferred to a universal tube containing 10 ml of RVS
on to Brilliant Green (BG) agar (CM0329; Oxoid), incubated at 37°C broth before incubation aerobically overnight at 41·5°C. If no growth
for 24 hours and examined for typical Salmonella colonies. Representative was recorded on the directly inoculated BG agar, a loopful (10 µl) of
swabs were also taken throughout the course of the vaccine trials. the inoculated RVS broth was streaked on to the surface of BG agar and
incubated at 37°C for 24 hours.
S Enteritidis challenge experiment Eggs were collected from each pair of birds, where possible, and cul-
When the birds were at the point of lay (17 weeks of age) they were tured individually for Salmonella. The outside of each egg was sterilised
re-housed in pairs in floor boxes. The birds in groups A, B, C and D by cleaning with 70 per cent ethanol, and the egg contents were then
were each separated at random into two equal-sized cohorts; the birds transferred aseptically to 90 ml of BPW and mixed thoroughly. The
in group E were separated into one group of 18, one group of 24 and one mixture was incubated at 37°C for 20 hours. Salmonellae were isolated
group of six. The birds in the first cohort were exposed to S Enteritidis following the selective enrichment procedure described above.
SE7D1 PT4 (an isolate from a caged layer flock) via water in fountain
drinkers: 3 x 102 cfu added to 250 ml of water in the base of the drinker Statistical analysis
(the ‘low dose’) for 24 hours. The birds in the second cohort were The proportions of Salmonella-positive birds in each of the experimental
exposed to 3 x 108 cfu of the same strain (the ‘high dose’), administered groups were compared using Pearson’s chi-squared test. All statistical
in the same way as the low dose challenge. Water consumption data for analyses were carried out using SPSS v 14.0.
Hy-Line Brown pullets, provided by the supplier, indicated that at 17
weeks of age each bird would normally consume approximately 180 ml Results
over a 24-hour period; therefore, each pair of birds would be expected The results of caecal colonisation for the Salmonella challenge studies are
to drink the whole 250 ml volume of water between them. As such, shown in Table 1. All birds were Salmonella-free before the experimental
assuming each bird drank 125 ml of the water, each would consume challenge. For the low Salmonella suspension (1·5 x 102 cfu), only the
approximately 1·5 x 102 cfu in the low dose challenge or 1·5 x 108 cfu in groups of birds vaccinated with LV1 or KV contained significantly fewer
the high dose challenge. The third group of six unvaccinated birds from S Enteritidis-positive birds than the unvaccinated group (P<0·018 and
group E remained unchallenged as a control for any salmonellae natu- P<0·044, respectively) (Table 2). Between the vaccinated groups, the
rally present in the environment. Following exposure, the birds were only significant difference was that birds vaccinated with LV1 had fewer
housed in pairs for 10 days before being euthanased. Salmonella-positive caeca than those vaccinated with LV2 (P=0·04)
(Table 2).
Enumeration of salmonellae The vaccinated groups were more difficult to differentiate following
Following dissection, the caecal contents from each bird were individu- challenge with the high (1·5 x 108 cfu) Salmonella suspension in terms
ally weighed and diluted to approximately 1:10 in buffered peptone of the absolute number of positive birds, although all vaccinated groups
water (BPW) (CM0509; Oxoid), thoroughly resuspended, and serial contained fewer Salmonella-positive birds than the unvaccinated group.
10-fold dilutions in 10 ml volumes of BPW were made down to 10–3. The proportion of Salmonella-positive birds was significantly lower in the
Duplicate volumes of 50 µl from each dilution were spread-plated on to groups vaccinated with LV1 (P<0·01) and KV (P<0·027) compared with
modified BG agar and incubated at 37°C for 24 hours before enumera- the unvaccinated group. There were no significant differences between
tion. All the dilutions in BPW were also incubated at 37°C for 20 hours. group B (vaccinated with LV2), group C (LV2+LV3) and the unvac-
Following incubation, 200 µl of each dilution of caecal contents was cinated group E when comparing the percentage of bird caecal contents
used to inoculate Diasalm medium (LAB537; LabM). These plates were testing positive for Salmonella. However, there were no significant dif-
incubated at 41·5°C for 24 to 48 hours. Presumptive Salmonella growth ferences between the four vaccinated groups. Group B (LV2) contained
after either 24 or 48 hours’ incubation was harvested and streaked onto a significantly lower proportion of birds for which Salmonella could
Rambach agar (1.07500.001; Merck) and incubated at 37°C for 24 be directly counted than the unvaccinated group (P<0·026), groups
hours before examination for typical Salmonella colonies. Presumptive A (LV1) and C (LV2+LV3) did not. Group D (vaccinated with KV)
Salmonella colonies on Rambach agar were confirmed by agglutination also contained a significantly lower proportion of birds that harboured
of poly-O, poly-H and O9 antisera (ProLab Diagnostics). directly countable numbers of Salmonellae in their caeca compared with

the VETERINARY RECORD | October 24, 2009

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TABLE 2: Results of chi-squared tests performed on the proportion of
Salmonella-positive birds in groups vaccinated with different vaccines
or left unvaccinated (control), following challenge with a low dose
(1·5 x 102 cfu) or a high dose (1·5 x 108 cfu) of Salmonella Enteritidis
Chi-squared value (P value)
LV1 LV2 LV3 KV Control*
Low challenge dose
LV1 4·21 (0·040) 1·18 (0·287) 0·15* (0·700) 5·6 (0·018)
LV2 0·45 (0·504) 2·86 (0·091) 0·11 (0·735)
LV2+LV3 0·50 (0·479) 1·78 (0·182)
KV 4·05 (0·044)
High challenge dose
LV1 1·6 (0·206) 0·64 (0·423) 0·15 (0·700) 7·57 (0·010)
LV2 0·23 (0·630) 0·371 (0·329) 0·360 (0·500)
LV2+LV3 0·18 (0·674) 1·13 (0·301)
KV 5·89 (0·027)
* One-tailed P value
KV Killed vaccine, LV Live vaccine
both the unvaccinated birds (P<0·0001) and the group vaccinated with
LV1 (P<0·003).
None of the 192 liver swabs or egg samples (20 samples per group (A
to E), 100 samples in total) tested positive for Salmonella in either the
vaccinated or unvaccinated groups.
Discussion
The aim of this study was to assess the protective effect of vaccination on
point-of-lay pullets when challenged with a field strain of S Enteritidis
under simulated commercial conditions. A lower percentage of point-
of-lay pullets vaccinated with KV or LV1 were colonised with Salmonella
following the low challenge, compared with unvaccinated birds. In addi-
tion, pullets vaccinated with LV1 were less likely to harbour Salmonella
than those given LV2 (P=0·04). However, there were no significant
differences in the total number of positive birds between the groups
given the different vaccines following the high Salmonella challenge.
Despite this, the most notable difference between the vaccinated and
unvaccinated groups following the high challenge was the level of cae-
cal colonisation. The group vaccinated with KV contained the smallest
number of birds with directly countable levels of Salmonella in their
caeca (≥102 cfu/g) compared with the unvaccinated group, followed by
the groups vaccinated with LV1 and LV3. Vaccination is one of several
factors that can influence the numbers of Salmonella recovered from the
caecal contents, and as such these data should be interpreted with cau-
tion. However, on layer farms harbouring few, if any, Salmonellae in the
birds’ environment, it is possible that vaccination could help to reduce
the overall numbers of Salmonella in a flock. In turn, this may reduce
infection rates and, when combined with effective cleaning, disinfection
and rodent control, help to prevent infection of subsequent flocks.
Vaccination has played a large part in reducing the contamination of
eggs in the UK (Cogan and Humphrey 2003). The S Enteritidis vaccines
currently available for the UK poultry industry are designed to provide
protection against infection from a low Salmonella challenge brought on
to the farm, for example, in feed, by wildlife or contaminated vehicles
or equipment, or a low level of residual contamination from a previous
Salmonella-positive flock. If Salmonella is introduced into a layer house
it can persist for prolonged periods, potentially resulting in the infec-
tion of successive flocks (Van de Giessen and others 1994). Sources of
Salmonella infection for newly placed pullets may include the contami-
nated house itself, rodents and others pests, or contaminated water and
feed (Gast 2005). Carry-over of infection from one flock to the next is
a common occurrence in UK laying houses, particularly in cage houses
(Carrique-Mas and others 2009), and has been observed in conjunc-
tion with poor cleaning and disinfection and inadequate rodent control.
Additionally, vertical transmission of some Salmonella strains is known
to occur from infected breeder flocks or as a result of contamination in
the hatchery (Methner and others 1995, Berchieri and others 2001).
This is a particularly important control point, as newly hatched chicks
are highly susceptible to colonisation of the intestinal tract with bacteria
(Duchet-Suchaux and others 1995, Gast and Benson 1996).
PAPERS
Despite the importance of vaccinating flocks against S Enteritidis,
vaccination may be insufficient to prevent colonisation in the face
of a high challenge. A high challenge may arise either when infected
rodents are present in the house (Carrique-Mas and others 2009) or fol-
lowing very deficient cleaning and disinfection between flocks (Wales
and others 2007). Thorough cleaning and disinfection of S Enteritidis-
positive laying houses is laborious and expensive, particularly in cage
houses (Wales and others 2006). These factors may allow high numbers
of Salmonella, harboured in organic material in the environment, to
overwhelm the protection provided by vaccines. Once a small number
of rodents or birds becomes infected, numbers of Salmonellae in the
environment will increase, increasing the likelihood of other vaccinated
birds in the flock becoming colonised.
As it is often difficult to remove all of the salmonellae in the envi-
ronment between flocks using conventional cleaning and disinfection
and pest control programmes, there is a strong case for improving the
effectiveness of vaccination programmes to protect against a higher
level of challenge. This short-term study has shown that vaccination
is effective in reducing the overall number of S Enteritidis-positive
birds and, in the case of KV and LV1, the level of colonisation. It is
uncertain, however, what the relative effect of these vaccines might
have been over the whole life of a flock under field conditions. The use
of vaccines and subsequent reduction in the numbers of S Enteritidis
detected in faecal samples may give a false impression that flocks are
negative for Salmonella when routine monitoring is performed. As
such, farmers may be unaware of the Salmonella status of their flocks
and so not be in an informed position to take further action to reduce
the carry-over of infection following flock depopulation. Therefore,
the use of such vaccines is likely to be most effective in controlling
Salmonella infection in laying flocks when used as part of a package
of measures including effective rodent control, terminal disinfection
and biosecurity.
Acknowledgements
This work was funded by Defra project OZ0325, with additional funding
from Intervet UK.
References
BARBOUR, E. K., FRERICHS, W. M., NABBUT, N. H., POSS, P. E. & BRINTON, M. K.
(1993) Evaluation of bacterins containing three predominant phage types of Salmonella
enteritidis for prevention of infection in egg-laying chickens. American Journal of Veterinary
Research 54, 1306-1309
BERCHIERI, A., WIGLEY, P., PAGE, K., MURPHY, C. K. & BARROW, P. A. (2001)
Further studies on vertical transmission and persistence of Salmonella enterica serovar
Enteritidis phage type 4 in chickens. Avian Pathology 30, 297-310
CARRIQUE-MAS, J. J., BRESLIN, M., SNOW, L., MCLAREN, I., SAYERS, A. R. &
DAVIES, R. H. (2009) Persistence and clearance of different Salmonella serovars in build-
ings housing laying hens. Epidemiology and Infection 137, 837-846
COGAN, T. A. & HUMPHREY, T. J. (2003) The rise and fall of Salmonella Enteritidis in
the UK. Journal of Applied Microbiology 94 (Suppl 1), 114-119
COYLE, E. F., PALMER, S. R., RIBEIRO, C. D., JONES, H. I., HOWARD, A. J., WARD,
L. & ROWE, B. (1988) Salmonella enteritidis phage type 4 infection: association with hen’s
eggs. Lancet 332, 1295-1297
CURTISS, R. & HASSAN, J. O. (1996) Nonrecombinant and recombinant aviru-
lent Salmonella vaccines for poultry. Veterinary Immunology and Immunopathology 54,
365-372
DAVIES, R. & BRESLIN, M. (2003) Effects of vaccination and other preventive meth-
ods for Salmonella Enteritidis on commercial laying chicken farms. Veterinary Record 153,
673-677
DAVISON, S., BENSON, C. E., HENZLER, D. J. & ECKROADE, R. J. (1999) Field obser-
vations with Salmonella enteritidis bacterins. Avian Diseases 43, 664-669
DE JONG, B. & EKDAHL, K. (2006) Human salmonellosis in travellers is highly correlated
to the prevalence of salmonella in laying hen flocks. Eurosurveillance. www.eurosurveil-
lance.org/ViewArticle.aspx?ArticleId=2993. Accessed May 13, 2009
DUCHET-SUCHAUX, M., LECHOPIER, P., MARLY, J., BERNARDET, P., DELAUNAY,
R. & PARDON, P. (1995) Quantification of experimental Salmonella enteritidis carrier
state in B13 leghorn chicks. Avian Diseases 39, 796-803
EFSA (2007a) The Community Summary Report on Trends and Sources of Zoonoses,
Zoonotic Agents, Antimicrobial resistance and Foodborne outbreaks in the European
Union in 2006. www.efsa.europa.eu/EFSA/efsa_locale-1178620753812_1178671312912.
htm. Accessed August 2, 2009
ESFA (2007b) Report of the Task Force on Zoonoses Data Collection on the analysis of
the baseline study on the prevalence of Salmonella in holdings of laying hen flocks of
Gallus gallus. www.efsa.europa.eu/EFSA/efsa_locale-1178620753812_1178620761896.
October 24, 2009 | the VETERINARY RECORD
 Downloaded from http://veterinaryrecord.bmj.com/ on May 19, 2015 - Published by group.bmj.com
PAPERS
htm. Accessed February 21, 2009
FEBERWEE, A., DE VRIES, T. S., HARTMAN, E. G., DE WIT, J. J., ELBERS, A. R. W. &
DE JONG, W. A. (2001) Vaccination against Salmonella enteritidis in Dutch commercial
layer flocks with a vaccine based on a live Salmonella gallinarum 9R strain: evaluation
of efficacy, safety, and performance of serologic Salmonella tests. Avian Diseases 45, 83-
91
GAST, R. K. (2005) Bacterial infection of eggs. In Food Safety Control in the Poultry
Industry. Ed G. Mead. Woodhead Publishing. pp 1-20
GAST, R. K. & BENSON, S. T. (1996) Intestinal colonization and organ invasion in chicks
experimentally infected with Salmonella enteritidis phage type 4 and other phage types iso-
lated from poultry in the United States. Avian Diseases 40, 853-857
GAST, R. K., STONE, H. D. & HOLT, P. S. (1993) Evaluation of the efficacy of oil-
emulsion bacterins for reducing fecal shedding of Salmonella enteritidis by laying hens. Avian
Diseases 37, 1085-1091
GILLESPIE, I. A., O’BRIEN, S. J., ADAK, G. K., WARD, L. R. & SMITH, H. R. (2005)
Foodborne general outbreaks of Salmonella Enteritidis phage type 4 infection, England and
Wales, 1992-2002: where are the risks? Epidemiology and Infection 133, 795-801
HOLT, P. S., STONE, H. D., GAST, R. K. & PORTER, R. E. (1996) Growth of Salmonella
enteritidis (SE) in egg contents from hens vaccinated with an SE bacterin. Food Microbiology
13, 417-426
HPA (2009a) Salmonella in humans (excluding S Typhi & S Paratyphi). Faecal and lower
gastrointestinal isolates reported to the Health Protection Agency Centre for Infections.
England and Wales, 1990-2008. www.hpa.org.uk/webw/HPAweb&HPAwebStandard/HP
Aweb_C/1195733760280?p=1191942172078. Accessed October 9, 2009
HPA (2009b) Joint Health Protection Agency and Food Standards Agency Press Release. 14
October 2004. Agencies step up action on Salmonella outbreaks linked to Spanish eggs.
www.hpa.org.uk/webw/HPAweb&HPAwebStandard/HPAweb_C/1195733760280?p=119
1942172078. Accessed January 16, 2009
LINDE, K., HAHN, I. & VIELITZ, E. (1996) Development of live Salmonella vaccines opti-
mally attenuated for chickens. Tierärztliche Umschau 51, 23-30 (In German)
METHNER, U., AL-SHABIBI, S. & MEYER, H. (1995) Experimental oral infection of
specific pathogen-free laying hens and cocks with Salmonella enteritidis strains. Journal of
the VETERINARY RECORD | October 24, 2009
Veterinary Medicine Series B 42, 459-469
MIYAMOTO, T., KITAOKA, D., WITHANAGE, G. S. K., FUKATA, T., SASAI, K. &
BABA, E. (1999) Evaluation of the efficacy of Salmonella enteritidis oil-emulsion bacterin
in an intravaginal challenge model in hens. Avian Diseases 43, 497-505
NAKAMURA, M., NAGAMINE, N., TAKAHASHI, T., SUZUKI, S. & SATO, S. (1994)
Evaluation of the efficacy of a bacterin against Salmonella enteritidis infection and the effect
of stress after vaccination. Avian Diseases 38, 717-724
PARKER, C., ASOKAN, K. & GUARD-PETTER, J. (2001) Egg contamination by
Salmonella serovar enteritidis following vaccination with Δ-aroA Salmonella serovar typh-
imurium. FEMS Microbiology Letters 195, 73-78
RABSCH, W., TSCHAPE, H. & BAUMLER, A. J. (2001) Non-typhoidal salmonellosis:
emerging problems. Microbes and Infection 3, 237-247
TIMMS, L. M., MARSHALL, R. N. & BRESLIN, M. F. (1990) Laboratory assessment of
protection given by an experimental Salmonella enteritidis PT4 inactivated, adjuvant vac-
cine. Veterinary Record 127, 611-614
VAN DE GIESSEN, A. W., AMENT, A. J. H. A. & NOTERMANS, S. H. W. (1994)
Intervention strategies for Salmonella enteritidis in poultry flocks: a basic approach.
International Journal of Food Microbiology 21, 145-154
WALES, A., BRESLIN, M. & DAVIES, R. (2006) Assessment of cleaning and disinfection
in Salmonella-contaminated poultry layer houses using qualitative and semi-quantitative
culture techniques. Veterinary Microbiology 116, 283-293
WALES, A., BRESLIN, M., CARTER, B., SAYERS, R. & DAVIES, R. (2007) A longitudi-
nal study of environmental Salmonella contamination in caged and free-range layer flocks.
Avian Pathology 36, 187-197
WOODWARD, M. J., GETTINBY, G., BRESLIN, M. F., CORKISH, J. D. & HOUGHTON,
S. (2002) The efficacy of Salenvac, a Salmonella enterica subsp enterica serotype
Enteritidis iron-restricted bacterin vaccine, in laying chickens. Avian Pathology 31, 383-
392
YAMANE, Y., LEONARD, J. D., KOBATAKE, R., AWAMURA, N., TOYOTA, Y.,
OHTA, H., OTSUKI, K. & INOUE, T. (2000) A case study on Salmonella enteritidis (SE)
origin at three egg-laying farms and its control with an S enteritidis bacterin. Avian Diseases
44, 519-526
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Salmonella colonisation of laying hens

following vaccination with killed and live
attenuated commercial Salmonella vaccines
R. J. Atterbury, V. M. Allen, J. J. Carrique-Mas and R. H. Davies

Veterinary Record 2009 165: 493-496

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