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Salmonella
the amount tested is usually governed by practical
considerations. In certain circumstances, however,
it may be necessary to test unusually large
sample amounts. During the investigation of baby
fooa-implicated in 3n Australia-wide outbreak of
infant salmonellosis, it was shown that sample
amounts of between 400-600 g per kg container
had to be examined before the causative
organism, Salmonella Bredeney, could be isolated
(104). In determining the quantity of sample to be
tested, consideration should be given to the
nature of the product, tnp pyppcted I@U€dftf
contamination, the probRnlp di"triblltion of
salmonellae within the product, and the purpos.e
of the examination, i.e. routine or invpstigRtion
~ The sample size and the number of sample
units (sub-samples) to be tested in the routine
examination of foods for salmonellae is usually
defined by a sampling plan. The establishment of
statistically based sampling plans for different
food categories is discussed in Chapter 3. These
sampling plans sometimes require the
examination of a large number of sample units to
attain a high probability of safety. For high risk
foods such as infant formulae, it has been
recommended that 6,.2individual 25 g sub-samples
should be tested with negative results (205). The
US Food and Drug Administration (FDA)
recommend sampling to the following plan
according to the nature of the food (19):
Category I. Foods that would not normally
be subjected to a process lethal to Salmonella
between the time of sampling and consumption
and are intended for consumption by the aged, the
infirm and infants (60 analytical units);
Category Il. Foods that would not normally
be subjected to a process lethal to Salmonella
between the time of sampling and consumption
(30 analytical units);
Category Ill. Foods that would normally be
subjected to a process lethal to Salmonella
between the time of sampling and consumption
(15 analytical units).
In most laboratories, routine examination of
such large sample numbers is not economically
feasible. The compositing or pooling of multiple
samples, however, provides a more practical
approach to Salmonella control: Huhtanen et al.
(185) examined 25 meat and bone meal samples
for salmonellae, comparing a si~ sample
with ten 30 g samples, and found that there was
little difference in isolation efficiency. Silliker and
Gabis (342) compared the efficiency of testing
sixty 30 g, fifteen 100 g and three 500 g sub-
samples for the recovery of salmonellae from
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naturally contaminated dried foods such as
chocolate bars, egg products, pepper, desiccated
coconut and meat and fish meals. Their results
demonstrated that salmonellae could be detected
with equal accuracy and with the same statistical
reliability using any of these sample pools.
Similar results were obtained in a subsequent
study of high moisture foods such as meat, poultry
and eggs (343). Compositing of samples is now
commonly used in many laboratories throughout
the world and is particularly advantageous for
foods where positive results are relatively rare. In
the laboratories of the FDA, 25 g sub-samples
may be composited to a maximum size of 375 g
(19). In Australia 375 g composite samples are
routinely used for many foods, especially dried
milk powders and AS 1766.2.5-1991 allows pooling
of up to 15 samples of the same batch of product.
Although sample units are usually measured
by mass, for some foods it may be more
appropriate to examine the external surface of the
product only. In this case test samples can be
obtained by excising, swabbing or washing
(rinsing) a defined surface area.
These sampling procedures are particularly
applicable to poultry, fish and some types of meat,
where salmonellae are unlikely to contaminate
the unexposed tissues. Excision of tissues
followed by maceration can provide precise data
on levels of surface contamination, but is not
generally favoured for routine work because of the
resultant downgrading of products. While surface
swabbing is often preferred because of the ease
and speed of manipulation, the level of recovery is
frequently inconsistent and incomplete. The
'whole bird rinse' technique described by
Surkiewicz et al. (364) had been widely
recommended as an effective, non-destructive and
practical method for the routine testing of poultry
for salmonellae (64, 96, 203, 290, 396). The
procedure involves rinsing both the external and
internal surfaces of the carcass in an appropriate
volume of a suitable diluent for 1-2 min using a
sterile plastic bag. The sensitivity of this method
has been shown to be related to the volume of
rinse fluid examined (73, 364) but rinse volumes
of 300, 500 and 1000 mL provide essentially equal
recovery efficiency (44). It has also been shown
that the sensitivity of the technique may be
improved by incubating the entire carcase in the
rinse solution (73). This latter procedure,
however, has not gained acceptance because ofthe
need for additional incubation space and because
the product is destroyed. As a general method for
examining food surfaces, a rinse technique is
probably suitable for most purposes. In Australia,
the rinse technique has been accepted as a
standard reference method for examining poultry
for salmonellae (358).
Specific discussion of the issues arising from
non-selective pre-enrichment and selective
233