BIOSECURITY AND VACCINATION IN
ERADICATING SALMONELLA ENTERITIDIS
Keith Gooderham, Poultry Health Services
10th September 1997
In 1985 we began to see a disease problem in
young chicks. Clinically they appeared as birds with
"Stunting and Runting " Disease.By about 10 days of age
some chicks had stopped growing and had distended abdomens.
Post-mortem examination revealed a persistent,inspissated yolk
sac and usually pericarditis. Salmonella enteritidis could
readily be grown on direct culture on solid medium (Blood
Agar,McConkeys Agar,Brilliant Green Agar).
This problem was widespread geographically, under different
ownership, in different breeds of chickens both broiler and layer
strains, as well as to a limited extent in turkeys and ducks.
Whilst, to my knowledge, the origin of the salmonella has not
been shown, I believe it originated in a large consignment of a
single feed ingredient. By the time the problem was recognised, a
secondary wave of the organism was being spread in the feed in
Poultry Offal Meal.
A junior health minister of the government was outspoken in
stating that all eggs being produced were from
salmonella-infected flocks. This was far from true, but public
concern was sufficient to result in legislation for testing all
flocks (breeding and laying ) and for slaughter of positive
flocks. The situation really was that caged egg layers were
seldom infected.
Broiler breeder flocks were frequently infected and my estimation
in 1989 was that 10% of broiler parent flocks carried the
organism.The same probably applied to layer parent flocks as well
as barn-egg and free-range layer flocks. Much microbiological
testing was carried out and somewhat later,serological testing
(when the ELISA kit became available ).
The main effort in my laboratory was directed at broiler parent
flocks. Much attention was focussed on feed mills and feed :
Poultry Offal Meal was no longer used,
Regular testing of other feed ingredients was increased,
Ministry compulsory testing of
home-produced and imported feed ingredients of animal
origin was introduced.
Flock testing over many flocks and many months showed :
1)Day-old parents did not carry S.enteritidis,
2)Flocks seldom became infected during rear to 18 weeks,
3)Flocks frequently became positive on the laying farm, usually either before 30 weeks of age or after 50 weeks of age,
4)Cleaning and disinfection of positive laying farms was frequently inadequate for preventing the next flock from becoming infected,
5)Hatcheries became contaminated due to receiving hatching eggs from positive flocks,
6)More flocks became infected as a result of spread from positive farms and hatcheries,
7)Infected mice on laying farms were
probably the most important source of
persistent infection.
There were so many positive flocks detected that
the Government Veterinary Department relaxed the slaughter
policy. A flock might still be slaughtered, but treatment of the
flock or its progeny were offered as alternatives. It was the
considerable effort put into eliminating the salmonella that
allowed us to learn its characteristics and pattern of spread.
All aspects of biosecurity were considered in detail and, where
possible, improved. Cleaning and disinfection of farms between
flocks was improved. Rodent control, particularly in preventing
their entry into livestock houses was
improved. As a result of all these measures, the incidence of
flocks infected with S.enteritidis diminished. Interestingly, the
cultural isolation of S. typhimurium and other, less invasive,
salmonellae were reduced to an even greater degree.
Competitive Exclusion.
The technique developed by Nurmi in Finland
involves supplying a range of cultured bacteria to the bird. The
idea is that these organisms establish themselves in the
intestine of the bird and compete with salmonella for sites at
which to multiply. Experience is varied. My own experience is
that it can be extremely effective against horizontal
transmission but is limited in its efficacy when transmission is
vertical. The value of competitive exclusion should not be
ignored but is only one part of the attempted reduction in
numbers of salmonella present. Of all the salmonellae which can
infect the chicken, S. enteritidis appears to be the
most resistent to control by the technique of competitive
exclusion.
Despite all efforts, S. enteritidis remained on certain
farm premises. Some had experienced 4 successive flocks becoming
infected, thus being "positive" for some 4 years. Some
Companies had greater problems of persistently contaminated
premises than others.
Vaccination
It was at a stage when such positive premises
were less than 5% of the total that a vaccine became available
for trial purposes. This was an inactivated vaccine administered
by injection as two doses given 4 weeks apart.
Initial government trials had been in commercial
layers, the vaccine being given at day-old and 4 weeks of
age.These were day-old to end of lay trials with groups being
taken at intervals for challenge studies.They were considered a
success. As broiler parents were not usually found to be
challenged during rear, and to speed up the time when vaccinated
birds were in lay, it was agreed to vaccinate at 12 and 16 weeks
of age. The plan was to vaccinate all flocks sending eggs to a
single hatchery so that eventually no eggs came from an
unvaccinated flock.This would take about a year from when the
vaccine was first available in March 1994.
Our experience was that the vaccine helped in the control of the
infection. Of the first 100 flocks vaccinated, 3 became infected
and transmitted the infection in their eggs. All 3 flocks were on
farms on which the previous flock had been positive. The next
cycle of vaccinated flocks on these premises remained negative.
Microbiology:
Culture of samples for salmonella is the most sensitive way of
detecting infected flocks. Some specific samples are required by
law. In addition to these we also test litter samples from flock
farms on a regular basis (once monthly).
This is a particularly sensitive test. With suspect positive eggs
in the hatchery,the most sensitive sample is to culture from
early dead embryos.These can be collected by candling at 10 days
of incubation. It is important when examining cultures to realise
that the appearance of colonies on the culture plate may not be
typical. It is possible to discard as negative what really is a
positive isolate.
Serology:
When the ELISA testing kit for S. enteritidis became available,
flocks were regularly tested to help identify positive flocks.
Results were not entirely accurate, with false positive
individuals appearing in negative flocks. Challenges from E.coli
could result in some sera "positive" to S.enteritidis.
A view of the flock had to be taken to determine whether it was
likely to be seroconverting. Further sera could be examined after
a few weeks. More importantly, however, a microbiological
examination of the flock should be made to confirm its salmonella
status.
As an example, a summary was made of all flocks tested for two
companies (A and B) each with several flock farms.
Company A had some positive flocks (by culture).
Company B had no positive flocks.
Figure 1. % Prevalence of positive sera according
to S/P reporting band
The scatter of mean flock titres against age for
the two companies is shown in the slides.
Figure 2, Company A : Mean S/P Bands according to flock age
Figure 3. Company B : Mean S/P bands according to flock age
Note the absence of positive titres during the
rearing period. In other companies practicing day-old-to-death
flock farm management, salmonella challenge was frequently very
early in rear. As flocks became vaccinated, serology was no
longer of value in identifying positive flocks.
Serological response to vaccination was, however, monitored.
Age of Vaccination:
As all flocks became vaccinated, it was decided to reduce the age
of vaccination. For convenience one company decided to vaccinate
at 2 and 6 weeks of age.
This was considered acceptable as the initial studies had shown
protection from a day-old and 4-week vaccination programme. It
became obvious that the serological response to the early
vaccination was less. Was this a problem, indicating lack of
protection, or was this useful in protecting the flock but
allowing recognition of the difference between vaccinated and
challenged flocks? Two early vaccinated flocks became positively
infected and egg-transmitted S. enteritidis. It was therefore
decided to revert back to vaccinating later, this time at 11 and
15 weeks of age.
Vaccination using the inactivated S.enteritidis vaccine
results in a serological response which will interfere with the
Pullorum Test. As indicated earlier, any positive serology should
be confirmed bacteriologically. Use of the vaccine together with
other S. enteritidis control measures would be a far
more effective way of controlling Pullorum Disease than using the
Pullorum Rapid Plate Test.
Vaccination of a Known S. enteritidis
Positive Flock.
A broiler parent chicken flock in two houses was found to be
infected and was transmitting S. enteritidis to its progeny. It
was decided to vaccinate one house and to treat the other with
Enrofloxacin.
Egg transmission was somewhat reduced.
At the end of lay 32 birds were culled from each house (8 birds
from each of the 4 pens in each house).
Organs were collected and cultured in pools of 4 . The organs
cultured were; liver +gall bladder, intestine including caecal
tonsils, oviduct, ovary.
Of the 32 pools of organs cultured from each house, there were 8
positive from each house. The positive organ pools were identical
for each house being; liver,1; intestines,2; oviduct,2; ovary,3.
It was concluded that the vaccination of an already positive
flock did not help in the control of S. enteritidis.
Opinion:
It is my opinion that the vaccine is an aid to controlling S.
enteritidis infection. It would not have worked against the
wealth of challenge that existed from 1985 to 1991. Strict
attention to biosecurity of farms, rodent control and cleaning
and disinfection programmes were vital in reducing the infections
to manageable proportions. These remain important. Also important
is the continued vigilance in the production of feed. Sourcing of
feed ingredients (which are still showing a level of 2% positive
samples, but rarely S. enteritidis), heat treatment of all feed
production and avoidance of recontamination of finished feed are
necessary.
HISTORY:
1984: First appearance of S. enteritidis PT4
1985: Disease in broiler chickens from egg-transmitted S.
enteritidis
1985-1988: Increasing infections in broiler parent flocks (10%)
1988 (Dec): Junior Health Minister speaks of problem
1989 (Mar): Legislation.Compulsory testing and slaughter
1989-1994: Attempts to eradicate not totally effective Repeatedly
positive farms identified Hatcheries repeatedly contaminated
1994 (Mar): Flock vaccination started
1994-1996: Reduction of positive farms Feed still a potential
source of Salmonella
1996: No positive flocks remain (Clients of Poultry Health
Services/Hillsdown Veterinary Services)
1997 (Sept): S. enteritidis cultured from the litter of
one flock. Enrofloxacin given. Egg transmission has not occurred