Campy Lob Act Ere Vibrio Ra
Campy Lob Act Ere Vibrio Ra
Campy Lob Act Ere Vibrio Ra
of Campylobacter spp.
in broiler chickens
FAO
FOOD AND
NUTRITION
PAPER
Nations
Bangkok, Thailand
5-9 August 2002
WORLD
HEALTH
ORGANIZATION
FOOD
SAFETY
CONSULTATIONS
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
ii
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
ACKNOWLEDGEMENTS
The Food and Agriculture Organization of the United Nations and the World Health Organization
would like to express their appreciation to the expert drafting groups (see Annex 2) for the time
and effort which they dedicated to the preparation of thorough and extensive technical documents
on risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafoods. The
deliberations of this expert consultation were based on these documents.
iii
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
iv
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
Contents
INTRODUCTION........................................................................................................................................1
BACKGROUND...........................................................................................................................................1
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
RECOMMENDATIONS ...........................................................................................................................47
vi
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
List of Abbreviations
CAC
CCFFP
CCFH
CFU
CT
ctx
FAO
FDA-VPRA
g
GHP
GMP
h
ISSC
MPN
PCR
ppt
RAP
TDH
tdh
TLH
TRH
trh
USFDA
WHO
vii
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
viii
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
1 Introduction
The Food and Agriculture Organization of the United Nations (FAO) and the World Health
Organization (WHO) convened an expert consultation on Risk assessment of Campylobacter spp. in
broiler chickens and Vibrio spp. in seafood in the FAO Regional Office for Asia and the Pacific
(RAP), Bangkok, Thailand on 5 - 9 August 2002. The list of participants is presented in Annex 1.
Mr Dong Qingsong, FAO Deputy Regional Representative for Asia and the Pacific and
Officer-in-charge, RAP, opened the meeting on behalf of the two sponsoring organizations. In
welcoming the participants Mr Qingsong noted the increasing significance of microbiological hazards
in relation to food safety. He noted that international trade had amplified the opportunity for these
hazards to be disseminated from the original point of production to locations thousands of miles away,
thereby permitting such food safety hazards to impact on public health and trade in more than one
country. Mr Qingsong observed that this underlined the need to first consider microbiological hazards
at the international level and provide the means by which they can then be addressed at regional and
national levels. He highlighted the commitment of FAO and WHO to provide a neutral international
forum to consider new approaches to achieving food safety, and in particular to address
microbiological risk assessment.
As the meeting was convened in Asia, Mr Qingsong also highlighted the importance of the
work on microbiological risk assessment for this region. He noted that seafood and poultry are
important products in both domestic and international trade and provide a valuable contribution to the
economy of the region. He also noted the concerns for public health raised by the microbiological
hazards associated with these foods. Therefore, in conclusion he requested the expert consultation to
consider, in particular, the practical application of their work so that the output would be of value to a
range of end users and to countries in different stages of development.
The consultation appointed Dr Serv Notermans (the Netherlands) as chairperson of the
consultation and Ms Dorothy-Jean McCoubrey (New Zealand) as rapporteur. Dr Notermans also
served as chairperson of the campylobacter working group and Prof. Diane Newell (United Kingdom)
served as rapporteur. Dr Mark Tamplin (United States of America) served as chairperson of the
working group on vibrio and Dr Ron Lee (United Kingdom) as rapporteur of that group.
2 Background
Risk assessment of microbiological hazards in foods has been identified as a priority area of
work for the Codex Alimentarius Commission (CAC). At its 32nd session the Codex Committee on
Food Hygiene (CCFH) identified a list of pathogen-commodity combinations for which it requires
expert risk assessment advice. In response, FAO and WHO jointly launched a programme of work
with the objective of providing expert advice on risk assessment of microbiological hazards in foods
to their member countries and to the CAC.
Dr. Hajime Toyofuku, WHO, and Dr. Sarah Cahill, FAO provided participants with an
overview of the joint FAO/WHO activities on microbiological risk assessment. In their presentation,
they also highlighted the objectives and expected outcomes of the current meeting.
The FAO/WHO programme of activities on microbiological risk assessment aims to serve
two customers - the CAC and the FAO and WHO member countries. The CAC, and in particular the
CCFH, needs sound scientific advice as a basis for the development of guidelines and
recommendations as well as the answers to specific risk management questions on certain pathogencommodity combinations. Member countries on the other hand need specific risk assessment tools to
use in conducting their own assessments and, if possible, some modules that can be directly applied in
a national risk assessment.
To implement this programme of work, FAO and WHO are convening a series of joint expert
consultations. To date three expert consultations have been held. The first of these was held on 17 - 21
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
July 20001 and the second on 30 April - 4 May 20012. Both of these expert consultations addressed
risk assessment of Salmonella spp. in broiler chickens and eggs and Listeria monocytogenes in readyto-eat foods. In March 2001, FAO and WHO initiated risk assessment work on the two pathogencommodity combinations being considered in this expert consultation. Two ad hoc expert drafting
groups were established to examine the available relevant information on Campylobacter spp. in
broiler chickens and Vibrio spp. in seafood and prepare documentation on both the exposure
assessment and hazard characterization steps of the risk assessment. These documents were reviewed
and evaluated by a joint expert consultation convened on 23 - 27 July 20013.
In October 2001, the report of that expert consultation was delivered to CCFH. Additional
risk management guidance was sought from the Committee in relation to the finalization of the risk
assessments. In response to this, the Committee established risk management drafting groups to
consider the approach it could take towards providing guidance on managing problems associated
with Campylobacter spp. in broiler chickens and Vibrio spp. in seafood. A limited amount of
feedback was received; therefore the risk assessment drafting groups, FAO and WHO met early in
2002 to consider how to complete the risk assessment work. Both risk assessments have been
progressed towards completion, although to varying extents. The risk assessment documents that have
been developed to date were reviewed and evaluated by the expert consultation.
The purpose of this report is to present the summary of the draft documents on risk
assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood as well as the
discussions and recommendations of the expert consultation.
To critically review the documents prepared by the ad hoc expert drafting groups giving
particular attention to;
the risk characterization component and the manner in which the risk assessment
outputs were generated;
To provide scientific advice to FAO and WHO member countries on the risk assessment of
Vibrio spp. in seafood and Campylobacter spp. in broiler chickens based on the available
documentation and the discussions during the expert consultation.
FAO. 2000. Joint FAO/WHO Expert Consultation on Risk Assessment of Microbiological Hazards in Foods, FAO
Headquarters, Rome, Italy, 17 - 20 July 2000. FAO Food and Nutrition Paper 71.
http://www.fao.org/es/ESN/food/risk_mra_campylobacter_en.stm / http://www.who.int/fsf/Micro/index.htm
2
FAO. 2001. Joint FAO/WHO Expert Consultation on Risk Assessment of Microbiological Hazards in Foods: Risk
characterization of Salmonella spp. in eggs and broiler chickens and Listeria monocytogenes in ready-to-eat foods, FAO
Headquarters, Rome, Italy, 30 April - 4 May 2001. FAO Food and Nutrition Paper 72.
http://www.fao.org/es/ESN/food/risk_mra_salmonella_en.stm / http://www.who.int/fsf/Micro/index.htm
3
WHO. 2001. Report of the Joint FAO/WHO Expert Consultation on Risk Assessment of Microbiological Hazards in
Foods; Hazard identification, exposure assessment and hazard characterization of Campylobacter spp. in broiler chickens
and Vibrio spp. in seafood. WHO Headquarters, Geneva, Switzerland 23 - 27 July 2001. WHO 2001.
http://www.fao.org/es/ESN/food/risk_mra_campylobacter_en.stm / http://www.who.int/fsf/Micro/index.htm
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
5.1.3 Scope
The purpose of the work was to develop a risk assessment that attempts to understand how the
incidence of human campylobacteriosis is influenced by various factors during chicken rearing,
processing, distribution, retail storage, consumer handling, meal preparation and finally consumption.
A benefit of this approach is that it enables consideration of the broadest range of intervention
4
WHO. 2001. Report of the Joint FAO/WHO Expert Consultation on Risk Assessment of Microbiological Hazards in
Foods; Hazard identification, exposure assessment and hazard characterization of Campylobacter spp. in broiler chickens
and Vibrio spp. in seafood. WHO Headquarters, Geneva, Switzerland 23 - 27 July 2001. WHO 2001.
http://www.fao.org/es/ESN/food/risk_mra_campylobacter_en.stm / http://www.who.int/fsf/Micro/index.htm
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
strategies. The risk characterization estimates the probability of illness per serving of chicken
associated with the presence of thermophilic Campylobacter spp. on fresh and frozen whole broiler
chicken carcasses with the skin intact and which are cooked in the domestic kitchen for immediate
consumption. A schematic representation of the risk assessment is shown in Figure 5.1.
Dose
Response
Prevalence
Farm
&
Transport
Slaughter
&
Processing
Preparation
&
Consumption
RISK
Concentration
FIGURE 5.1: Schematic representation of the risk assessment of Campylobacter spp. in broiler
chickens.
5.1.4 Approach
5.1.4.1 Hazard identification
Thermophilic Campylobacter spp. are a leading cause of zoonotic enteric illness in most
developed countries (Friedman et al., 20005). Human cases are usually caused by C. jejuni, and to a
lesser extent, by Campylobacter coli. Information on the burden of human campylobacteriosis in
developing countries is more limited (Oberhelman & Taylor, 20006). Nevertheless, it is reported that
asymptomatic infection with C. jejuni and C. coli is frequent in adults. In children, under the age of
two, C. jejuni, C. coli and other Campylobacter spp. are all associated with enteric disease.
Campylobacter spp. may be transferred to humans by direct contact with contaminated
animals or animal carcasses or indirectly through the ingestion of contaminated food or water. The
principal reservoirs of thermophilic Campylobacter spp. are the alimentary tracts of wild and
domesticated mammals and birds. Campylobacter spp. are commonly found in poultry, cattle, pigs,
sheep, wild animals and birds, and in dogs and cats. Foodstuffs, including, poultry, beef, pork, other
meat products, raw milk and milk products, and, less frequently, fish and fish products, mussels and
fresh vegetables can also be contaminated (Jacobs-Reitsma, 20007). Findings of analytical
epidemiological studies are conflicting. Some have identified handling raw poultry and eating poultry
products as important risk factors for sporadic campylobacteriosis (Friedman, et al., 20005), however
others have found that contact in the home with such products is protective (Adak et al., 19958).
Friedman, C., Neimann, J., Wegener, H. & Tauxe, R. 2000. Epidemiology of Campylobacter jejuni infections in the United
States and other industrialized nations. In Campylobacter 2nd Edition, pp. 121-138. Edited by I. Nachamkin & M. J. Blaser.
Washington DC: ASM Press.
6
Oberhelman, R. & Taylor, D. 2000. Campylobacter infections in developing countries. In Campylobacter 2nd Edition, pp.
139-154. Edited by I. Nachamkin & M. Blaser. Washington DC: ASM Press.
7
Jacobs-Reitsma, W. 2000. Campylobacter in the food supply. In Campylobacter 2nd Ed., pp. 467-481. Edited by I.
Nachamkin & M. J. Blaser. Washington DC: ASM Press.
8
Adak, G. K., Cowden, J. M., Nicholas, S. & Evans, H. S. 1995. The Public Health Laboratory Service national case-control
study of primary indigenous sporadic cases of campylobacter infection. Epidemiology and Infection 115, 15-22.
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
Information for the hazard identification section was compiled from published literature and
from unpublished data submitted to FAO and WHO by public health agencies and other interested
parties.
5.1.4.2 Hazard characterization
Hazard characterization provides a description of the public health outcomes following
infection, including sequelae, pathogen characteristics influencing the organism's ability to elicit
infection and illness, host characteristics that influence the acquisition of infection, and food-related
factors that may affect the survival of C. jejuni in the human gastrointestinal tract. A dose-response
model was derived that mathematically describes the relationship between the numbers of organisms
that might be present in a food and consumed (the dose), and the human health outcome (the
response). In order to achieve this, human feeding trial data (Black et al., 19889) for two strains of
C jejuni were pooled and used to derive estimates for the probability of infection (colonization of the
gastrointestinal track without overt symptoms) as well as the probability of illness once infected.
Campylobacteriosis predominantly occurs as sporadic cases (Friedman, et al., 200010), and hence
dose-response data from outbreak investigations are essentially non-existent.
The probability of infection upon ingestion of a dose of C. jejuni was estimated with the
caveat that the data are from a feeding study involving healthy volunteers, and using a milk matrix
and a limited number of campylobacter strains. Whether the probability of infection and/or illness are
different for immune individuals or those individuals with an increased susceptibility to illness (e.g.
immunosuppressed, very young, etc.) compared to the volunteers could not be determined from the
feeding trial data. The probability of illness following infection was also estimated based upon these
investigations, and assumed to be dose-independent. Again, the impact of other factors, such as
susceptibility, on the probability of illness cannot be quantified due to a lack of adequate
epidemiological data and resolution to this level. The progression of the illness to more serious
outcomes and the development of some sequelae can be crudely estimated from the approximate
proportions reported in the literature, but these were not included in the present work. For the
purposes of this model it was assumed that C. coli has the same properties as C. jejuni.
5.1.4.3 Exposure assessment
The exposure model describes the stages Rearing and Transport, Slaughter and Processing,
and Preparation and Consumption (Figure 5.1). This modular approach estimates the prevalence and
concentration of thermophilic Campylobacter spp., and the changes in these associated with each
stage of processing, and refrigerated and frozen storage and consumer handling of the product.
Routes contributing to initial introduction of Campylobacter spp. into a poultry flock11 on the
farm have been identified in epidemiological studies. However, these remain poorly understood and
the phenomenon may be multi-factorial, and hence, colonization was modelled based on an
unspecified route of introduction. However, an epidemiology module has been created as a
supplement to the model, which allows evaluation of interventions aimed at changing the frequency
with which risk factors occur. As such risk factors are usually interdependent and may be country
specific, incorporation of this module into the current risk model was not appropriate: however, it
does illustrate how such information may be used in a specific situation.
The effects of cooling and freezing, and adding chlorine during water chilling, are evaluated
in the model. Effects of other processing treatments, e.g. lactic acid and irradiation, on reduction of
Campylobacter spp. contamination on poultry carcasses were not explicitly evaluated as the
quantitative data are lacking: however, if the level of reduction as a consequence of any type of
9
Black, R. E., Levine, M. M., Clements, M. L., Hughes, T. P. & Blaser, M. J. 1988. Experimental Campylobacter jejuni
infection in humans. Journal of Infectious Diseases 157, 472-9.
10
Friedman, C., Neimann, J., Wegener, H. & Tauxe, R. 2000. Epidemiology of Campylobacter jejuni infections in the
United States and other industrialized nations. In Campylobacter 2nd Edition, pp. 121-138. Edited by I. Nachamkin & M. J.
Blaser. Washington DC: ASM Press.
11
A flock is a group of hens of similar age that are managed and housed together.
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
processing modification, can be quantified for any treatment, then the impact on risk can be readily
evaluated within the risk model.
The home preparation module considered two routes of ingestion of viable organisms: via
consumption of contaminated, undercooked chicken meat, and via cross-contamination, from
contaminated raw poultry onto another food that is eaten without further cooking, or directly from the
hands (Figure 5.2). As a result of recommendations from the previous expert consultation12 the
protected areas cooking approach and the drip-fluid model approach for cross-contamination were
selected. The "protected areas" approach to modelling the cooking step considers that campylobacter
cells may be located in parts of the carcass that reach cooking temperature more slowly than other
locations. The drip-fluid model for cross-contamination relates to the spread of campylobacter cells
via drip-fluid from raw carcasses.
Raw Chicken
Cross contamination
Under cooking
Exposure
FIGURE 5.2: Schematic representation of the home preparation module.
5.1.4.4 Risk characterization
The risk characterization step integrates the information collected during the hazard
identification, hazard characterization, and exposure assessment steps to arrive at estimates of adverse
events that may arise due to consumption of chicken (Figure 5.1). This step links the probability and
magnitude of exposure to campylobacter associated with consumption of chicken to adverse outcomes
that might occur. The resulting risk is expressed as individual risk or the risk per serving of chicken.
Although this model does not address risk to a specific population, data on amounts of chicken
consumed can be incorporated into the model to arrive at estimates of the population-based risk.
The current model is unable to provide a central estimate of risk due to virtually unbounded
uncertainty13 on two key components of the model, namely the impact of undercooking and the
impact of cross-contamination. Since these processes are the ultimate determinant of the final
exposure of the consumer, the unbounded and unresolvable nature of the uncertainty undermines the
establishment of a central estimate of consumer risk. In addition, the model has not yet been applied
12
WHO. 2001. Report of the Joint FAO/WHO Expert Consultation on Risk Assessment of Microbiological Hazards in
Foods; Hazard identification, exposure assessment and hazard characterization of Campylobacter spp. in broiler chickens
and Vibrio spp. in seafood. WHO Headquarters, Geneva, Switzerland 23 - 27 July 2001. WHO 2001.
http://www.fao.org/es/ESN/food/risk_mra_campylobacter_en.stm / http://www.who.int/fsf/Micro/index.htm
13
As the upper and lower boundaries of the values describing the impact of undercooking and cross-contamination are
unknown the distribution of possible values is potentially infinite and therefore unbounded.
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
to explore the upper and lower bound estimates of risk that are suggested by the uncertainty.
However, the model still provides a useful means of studying potential exposure pathways and how
these contribute to the risk of illness posed by campylobacter associated with broiler chickens.
The risk characterization involves the following elements:
1) A baseline model
A baseline model was explored in detail, to clarify the input-output relations between the
different modules of the model and also to explore the credibility of the model. The baseline model
was defined as the processing of fresh carcasses from both positive and negative flocks (with an
overall flock prevalence of 80%), which are air chilled at the end of slaughter.
The model indicated that the external campylobacter load per chicken increased during
transport and evisceration, and decreased at the other processing steps studied, with an overall
reduction of the mean load from farm to fork of about 4 to 5 logs (Figure 5.3). The prevalence of
campylobacter-contaminated chickens from positive flocks appears to drop from 100% of live birds to
20% of chicken meat servings (Figure 5.4). For negative flocks, prevalence increases during transport,
defeathering and evisceration, indicating the effect of cross-contamination during processing.
Prevalence later drops to a value of about 3% of servings at the moment of consumption.
The correlation between the input and output of the models of the different processing steps
has been graphed to illustrate the variability in load per chicken along the process. These plots
indicate that the load on a chicken at the stages before evisceration appears to be a bad predictor for
the final load, and thus for the risk per serving of that particular chicken carcass.
2) Scenario analysis
In a scenario analysis, the effects of alterations to the baseline are studied by changing one or
two of the model parameters. By doing so the impact of uncertainty in parameter estimates can be
explored, and the potential of risk mitigation strategies can be evaluated.
Eight alternative scenarios were explored, to illustrate the ability of the model to investigate
the effect of changing specific model inputs. Each individual scenario involved a specific change in
one parameter, as follows:
Mitigation related scenarios
1. Between flock14 prevalence (80% prevalence reduced to 50%)
2. Within flock prevalence (100% prevalence reduced to 10%)
3. Scalding (hard scald (56-58C for 2-2.5 minutes)and soft scald (50-52 C for up to
3.5 minutes))
Alternate model assumption scenarios
4. Shorter freezing storage time range (1 day to 6 weeks storage time compared to 1 day
to 1 week storage time)
5. Undercooking (5C lower temperature in coolest spot in the chicken during cooking)
14
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
10
8
6
4
2
0
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FIGURE 5.3: Numbers of campylobacter cells per carcass during processing of fresh air-chilled
carcasses, from both positive and negative flocks. (Negative flocks get contaminated during
transport). Both the mean of the logs and the log of the means are given. (The differences between
these is as a result of the skewness15 of the distribution of values and the fact that 'zero'- values cannot
be incorporated in calculations of the mean of logs (which is therefore only about the positive
log mean of positive flocks
carcasses)). ____________
mean log of positive flocks
log mean of negative flocks
mean log of negative flocks
do
se
tra
ns
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rt
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ald
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0.9
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0.0
fa
rm
Prevalence
----------______ ______
x
------x------
FIGURE 5.4: The prevalence of campylobacter on fresh air-chilled broiler chicken carcasses coming
from flocks that were positive and negative for campylobacter at the farm level.
Flocks positive for campylobacter at the farm
Flocks negative for campylobacter at the farm
15
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
To further detail the model analysis, and to determine the effect of introducing different
mitigations, five series of simulations were run. Using the baseline model settings one of following
parameters was changed in each series of simulations:
flock prevalence;
campylobacter load on carcasses after transport;
campylobacter load in caeca;
campylobacter load in both caeca and on carcasses after transport;
campylobacter load on carcasses after evisceration.
A linear relationship between flock prevalence and probability of illness was found. Thus a
two-fold reduction in flock prevalence would result in a corresponding two-fold reduction in the
probability of illness. This effect is mainly caused by a reduction in the number of campylobacterpositive meals. Both external contamination and colonization need to be reduced concurrently to
achieve a substantial impact on risk. This is due to the potential for large numbers of campylobacter
cells contaminating the carcass as a result of damage to the viscera at evisceration, thus undermining
any benefits achieved in reducing the external numbers at an earlier processing step. Use of the model
has identified two key ways of achieving this reduction: 1) reducing colonization early enough to
impact external contamination and 2) intervening post-evisceration to reduce carcass contamination.
3. It was difficult to model cross-contamination in the home as a result of the lack of a clear
understanding of the pathways and lack of data quantifying the magnitude and frequency of crosscontamination events. Further improvement of this module and validation may be extremely
difficult given the complexity of cross-contamination, the many possible pathways by which it
can occur, and the variability in the behaviour of individuals in the kitchen.
4. Based on thermal inactivation calculations, it was difficult to reconcile the assumed importance of
undercooking as a cause of human exposure to campylobacter, if the contamination of broiler
carcasses with campylobacter is on the external surface of the carcass (or very close to the
surface). Resolution of this inconsistency requires the allocation of some amount of
contamination to sub-surface sites within the carcass where the temperature increases much more
slowly. While it is possible to demonstrate that campylobacter will, on occasion, be found in such
places, it is very difficult to quantify the frequency and extent of this particular mode of
contamination.
10
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
8. Assuming that cooking performance is independent of the chicken being fresh or frozen, frozen
chicken posed a lower risk via consumption than fresh chicken.
9. The washing-off effect associated with water chilling translated to water-chilled chickens posing a
lower risk than air-chilled chickens. However, there was uncertainty associated with the degree of
cross-contamination that occurs in the chill tank during water chilling that would have an impact
on this comparison and may be affected by the addition of chlorine to the chill water.
10. Undercooking was estimated to have a higher risk than cross-contamination using one set of
assumptions. The cooking and cross-contamination modules are based on plausible theoretical
constructs, but knowledge and data related to these two pathways are essentially unavailable.
Since the set of assumptions on which this comparison relied was only one, of many, plausible
sets, analysis of this component of the model remains inconclusive.
11. Unlike many other mitigation scenarios, there was very little uncertainty that reduction of the
between-flock prevalence of campylobacter would reduce any associated public health risk. A
linear relationship was found to exist between flock prevalence and probability of illness, i.e. a
two-fold reduction in flock prevalence would result in a corresponding two-fold reduction in the
probability of illness.
12. In order to meaningfully reduce the bacterial load on processed carcasses, interventions would be
required to address the bacterial load both internally and externally, since efforts directed at only
one of these contamination reservoirs can be readily undermined by high levels of contamination
from the other.
16
The process of elaborating FAO/WHO "Guidelines for incorporating quantitative risk assessment in the development of
microbiological food hygiene standards, guidelines and related texts" began at a consultation convened in Kiel, Germany on
18 - 22 March 2002. The draft guidelines are currently being finalized and will be available on the FAO and WHO webpages
at the end of 2002. http://www.fao.org/es/ESN/food/risk_mra_riskmanagement_en.stm / http://www.who.int/fsf/Micro/index.htm
11
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
assessment process. Data collection, one of the most important related activities, received particular
attention and focus was given to the main tasks that need to be undertaken.
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
5.2.1 Introduction:
The expert consultation highlighted the importance of including in the introduction of the
final risk assessment document a succinct, clear description of the history of this exercise: by whom it
was initiated; what reports have been produced, and when. The objectives of the risk assessment
should be more clearly stated, with reference to who suggested them. Objectives which have been
specifically excluded should also be listed. It would be useful to state whether each objective has been
achieved and if not, why not. This would facilitate a better understanding of the work and why it was
developed in such a manner. Descriptions of methods and approaches should not appear in this
section.
A concluding paragraph suggesting the steps to be taken following the successful conclusion
of this initiative would be beneficial to the end-users, while also stressing that no risk assessment
model is complete as long as data gaps exist.
13
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
All strains have the same colonization potential and persistency in the chicken gut:
with the current level of knowledge this is the only assumption that can be made, but
it may be untrue.
There are only three transmission scenarios.
Birds stay in their social clusters during harvesting. However, it is known that birds
move during harvesting so the social clusters are unlikely to be maintained.
Data deficiencies and recommendations for model improvements
It is evident that the reduction or elimination of campylobacter colonization in the flock is
extremely important. At the farm level there are limited strategies to achieve this aim. Approaches
include preventing flock exposure by biosecurity or reducing bird susceptibility to colonization by
measures such as vaccination or competitive exclusion treatment (Newell & Wagenaar, 200017). The
latter approaches are not yet available commercially and, therefore, biosecurity is the only strategy
currently feasible. A module to assess the relative importance of sources of colonization would be
extremely useful to risk managers, and such a module has been initiated. However, it was considered
by the risk assessment drafting group that there were at this time insufficient data on flock infection
sources for the use of such a module.
The part of the model dealing with the effect of catchers contaminating the exterior of birds
could, with some modification, be adapted to model the effects of thinning (the early removal of a
proportion of the birds) which is considered a major source of broiler colonization.
Using data on between-flock and within-flock prevalence of campylobacter the probability of
any random bird being campylobacter-positive can be estimated. The model also demonstrates that
the probability of colonization is dependant on age which is consistent with available data (Newell
and Wagenaar, 200017). Although the model can indicate the effect of various generic sources on
transmission it cannot, at this time, allow an assessment of the sources of exposure to provide targeted
strategies for intervention.
5.2.3.2 Processing
The main features of this module are:
A model which mimics the changes in level of contamination after scalding,
defeathering, evisceration and chilling.
A provision in the model to differentiate the effects of air and water chilling and the
use of super-chlorination.
An estimation of the prevalence of contaminated products
A provision to model the changes in level of contamination after storage at 4 oC or
frozen at -20 oC.
This part of the model assumes that:
Campylobacter do not grow outside the host gut. This is consistent with most
scientific evidence (Jacobs-Reitsma, 200018).
There are a finite number of sites of cross-contamination within a plant. There are
however many more sites of potential cross-contamination within a processing plant
other than scalding, defeathering and evisceration but these would appear to include
the most important sources (Mead 198919).
Data deficiencies and recommendations for model improvements
The poultry production industry has substantially improved hygiene control over the last 30
years. Many of the changes may have effects on the data entered into the model, for example the
17
Newell, D. G. & Wagenaar, J. A. 2000. Poultry infections and their control at the farm level. In Campylobacter, 2nd Ed.,
pp. 497-509. Edited by I. Nachamkin & M. J. Blaser. Washington DC: ASM Press.
18
Jacobs-Reitsma, W. 2000. Campylobacter in the food supply. In Campylobacter 2nd Ed., pp. 467-481. Edited by I.
Nachamkin & M. J. Blaser. Washington DC: ASM Press.
19
Mead G.C. 1989. Hygiene problems and control of process contamination. In: Processing of poultry (ed Mead G.C.)
Chapman Hall, London.pp 183-220.
14
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
introduction of multistage scalding and newer evisceration machinery which separates the viscera
from the carcass. If there are data or evidence that such changes have a significant effect on
campylobacter contamination of carcasses then appropriate changes to the model should be
considered.
Overall, the spread of contamination from the gut contents of positive birds will have the
greatest effect on the level of contamination on external bird surfaces. However, as the proportion of
negative flocks increases, the importance of cross-contamination becomes more apparent. Data is now
being collected for the level of contamination on carcasses from campylobacter-negative flocks. This
data should be available in the near future, and the expert consultation recommended that at that time
it should, if possible, be incorporated into the model.
The model should take account of the published and unpublished data showing that organisms
attached to the carcass surface are resistant to environmental effects. (Notermans and Kemplemaker,
197520; G. Mead, personal communication, 2002).
Only freezing and chilled storage were considered in the post-processing part of this module.
Recent anecdotal data indicates that modified atmosphere packaging has an impact on levels of
campylobacter contamination on poultry products. The expert consultation recommended that when
this data comes into the public domain it should, if possible, be incorporated into this module.
There is increasing evidence for variation in the survival of campylobacter strains during
processing (Newell et al., 200121). This may have a considerable effect on the model especially if the
ability of Campylobacter spp. to survive is associated with strain virulence. At this time there are no
simple methods for assessing the ability to survive but models for this property are available and data
is currently being accumulated. Once this data becomes available the expert consultation
recommended that it should, if possible, be incorporated into this module.
5.2.3.3 Consumer handling and cooking
The main features of this part of the model are:
Estimates of the proportion of loosely attached campylobacter cells associated with
poultry.
Estimates of the concentration of campylobacter cells in the drip-fluid from a random
wet chilled carcass.
A model of the effect of temperature, time of exposure and location at a protected site
on campylobacter survival.
This part of the model assumes that:
10-20% of campylobacter cells are located at protected sites. The expert consultation
considered this assumption to be unexpectedly high.
Data deficiencies and recommendations for model improvements
More information is required on consumer practices in domestic kitchens. Information is
needed on routes and modes of transmission of campylobacter within the kitchen. Factors affecting
campylobacter survival in the kitchen environment are also required.
20
Notermans S. and Kamplemaker E.H. 1975. Heat destruction of some bacterial strains attached to broiler skin. British
Poultry Science, 16: 351-361
21
Newell, D. G., Shreeve, J. E., Toszeghy, M., Domingue, G., Bull, S., Humphrey, T. & Mead, G. 2001. Changes in the
carriage of campylobacter strains by poultry carcasses during processing in abattoirs. Applied and Environmental
Microbiology. 67, 2636-40.
15
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
22
Oberhelman, R. & Taylor, D. 2000. Campylobacter infections in developing countries. In Campylobacter 2nd Edition, pp.
139-154. Edited by I. Nachamkin & M. Blaser. Washington DC: ASM Press.
23
Cawthraw, S. A., Lind, L., Kaijser, B. & Newell, D. G. 2000. Antibodies, directed towards Campylobacter jejuni antigens,
in sera from poultry abattoir workers. Clinical and Experimental Immunology 122, 55-60.
24
Newell, D. & Nachamkin, I. 1992. Immune responses directed against Campylobacter jejuni. In Campylobacter jejuni:
Current staus and future trends, pp. 201-206. Edited by I. Nachamkin, M. Blaser & L. Tompkins. Washington DC: ASM
Press.
16
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
The expert consultation identified the following research priorities, however, it is critical to
note that research priorities will vary depending on the specific risk management question the risk
assessment is being used to address.
A. Exposure assessment
Information about the influence of strain-specific variation on campylobacter survival on
poultry meat.
Information on consumer and retail practices in relation to the risk of transfer of
campylobacter from poultry products to kitchen surfaces and other foods.
Data on the routes of campylobacter colonization of broilers at the farm level so that farm
interventions can be appropriately targeted.
B. Hazard characterization
Additional data on dose-response.
Data on strain variability in relation to virulence and pathogenicity.
17
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
WHO. 2001. Joint FAO/WHO Expert Consultation on Risk Assessment of Microbiological Hazards in Foods; Hazard
identification, exposure assessment and hazard characterization of Campylobacter spp. in broiler chickens and Vibrio spp. in
seafood, WHO Headquarters, Geneva, Switzerland 213 - 27 July 2001. WHO 2001.
http://www.fao.org/es/ESN/food/risk_mra_campylobacter_en.stm / http://www.who.int/fsf/Micro/index.htm
26
Report of the thirty fourth session of the Codex Committee on Food Hygiene, Bangkok, Thailand, 8 - 13 October 2001
ALINORM 03/13 para. 77. http://www.codexalimentarius.net/reports.asp
18
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
Possible intervention measures should be given in the report to illustrate actions that could be
taken by risk managers.
Technical details given in the background information on production and processing need to be
updated.
27
Oliver, J. D., and Kaper, J.B. 1997. Vibrio Species. In M. P. Doyle, L. R. Beuchat, and T. J. Montville, eds. Food
Microbiology: Fundamentals and Frontiers, p228-264. Washington, D.C., ASM Press.
28
Dalsgaard, A. 1998. The occurrence of human pathogenic Vibrio spp. and Salmonella in aquaculture. International
Journal of Food Science and Technology, 33: 127-138.
29
Desmarchelier, P.M. 1997. Pathogenic Vibrios. In A.D. Hocking, G. Arnold, I. Jenson, K. Newton and P. Sutherland, eds.
Foodborne Microorganisms of Public Health Significance 5th Edition, p 285 -312. North Sydney, Australian Institute of
Food Science and Technology Inc.
30
Dalsgaard, A. Mller, N.F., Brin, B., Hoei, L. and Larsen, J.L. 1996. Chemical manifestation and epidemiology of Vibrio
vulnificus in Denmark (summer 1999). European Journal of Clinical Microbiology and Infectious diseases 15. 227 - 232.
19
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
6.1.2 Scope
The risk assessment work was undertaken on the following pathogen-commodity
combinations:
Vibrio parahaemolyticus in raw oysters consumed in Japan, New Zealand, Canada,
Australia and the United States of America.
Vibrio parahaemolyticus in finfish consumed raw.
Vibrio parahaemolyticus in bloody clams consumed in Thailand.
Vibrio vulnificus in raw oysters consumed in the United States of America.
Choleragenic Vibrio cholerae in warm-water shrimp in international trade.
TABLE 6.1: Vibrio spp. which cause, or are associated with, human infections (after Dalsgaard,
199831)
Occurrence in human clinical specimens*
Intestinal
V. cholerae O1 and O139
V. cholerae non-O1/non-O139
V. parahaemolyticus
V. fluvialis
V. furnissii
V. hollisae
V. mimicus
V. metschnikovii
V. vulnificus**
V. alginolyticus
V. carchariae
V. cincinnatiensis
V. damsela
++++
++
++++
++
++
++
++
+
+
-
Non-intestinal
+
++
+
+
+
+++
++
+
+
+
*The symbol (+) refers to the relative frequency of each organism in clinical specimens and (-) indicated that the organism
was not found
**The ability of V. vulnificus to cause gastro-intestinal disease remains to be confirmed
Dalsgaard, A. 1998. The occurrence of human pathogenic Vibrio spp. and Salmonella in aquaculture. International
Journal of Food Science and Technology, 33: 127-138.
20
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
development of a risk model. A critical component of the model was water temperature. Since high
water temperatures are a factor in several countries with significant oyster industries, FAO and WHO
decided to undertake a risk assessment on consumption of raw oysters in a number of different
countries. As well as generating an estimate of the number of annual illnesses, a further aim was to
assess the potential of the model developed in the United States of America to predict oyster-borne
V. parahaemolyticus illness from oysters grown in different regions and using different production
systems.
6.1.3.2
Scope
The risk assessment covers consumption of raw oysters in five countries: New Zealand,
Japan, Canada, Australia and the United States of America.
6.1.3.3 Hazard identification
V. parahaemolyticus has been recognized as a major cause of seafood-borne gastroenteritis in
Japan (Twedt, 198932; Ministry of Health, Labour and Welfare, Japan, 200033) and other Asian
countries. By contrast, in most countries outside of Asia, the reported incidence appears to be low,
perhaps reflecting a different mode of seafood consumption. Gastroenteritis caused by this organism
is almost exclusively associated with seafood consumed raw or inadequately cooked, or contaminated
after cooking. In the United States of America, prior to 1997, illness was most commonly associated
with crabs, oysters, shrimp and lobster (Twedt, 198932; Oliver and Kaper, 199734). Four
V. parahaemolyticus outbreaks associated with the consumption of raw oysters were reported in the
United States of America in 1997 and 1998 (DePaola et al., 200035). A new V. parahaemolyticus
clone of O3:K6 serotype emerged in Calcutta in 1996. It has spread throughout Asia and to the United
States of America elevating the status of V. parahaemolyticus to pandemic (Matsumoto et al., 200036).
In Australia, in 1990 and 1992, there were two outbreaks of gastroenteritis caused by
V. parahaemolyticus in chilled, cooked shrimps imported from Indonesia (Kraa, 199537) and there was
also a death in 1992 associated with the consumption of oysters.
6.1.3.4 Hazard characterization
This section focuses on evaluating the nature of adverse health effects associated with
V. parahaemolyticus in seafood and how to quantitatively assess the relationship between the
magnitude of the food-borne exposure and the likelihood of adverse effects occurring. It included the
elaboration of a dose-response curve. Infection by V. parahaemolyticus is characterized by an acute
gastroenteritis. Therefore, the end-point of the dose-response curve was defined as gastroenteritis.
A review of the literature was undertaken to identify and characterize the infectivity and
genetic factors of V. parahaemolyticus, which has both pathogenic and non-pathogenic forms based
on the presence of specific virulence genes: tdh (thermostable direct haemolysin gene) and trh (TDHrelated haemolysin gene). Relevant factors with respect to the host and food matrix have been
identified and where data are available may be incorporated into the model.
32
Twedt, R. M. 1989. Vibrio parahaemolyticus. In M. P. Doyle, ed. Foodborne Bacterial Pathogens, p543-568. New York,
Marcel Decker, Inc.
33
Ministry of Health, Labour and Welfare, Japan 2000. Statistics of Food Poisoning Japan in 2000.
34
Oliver, J. D., and Kaper, J.B. 1997. Vibrio Species. In M. P. Doyle, L. R. Beuchat, and T. J. Montville, eds. Food
Microbiology: Fundamentals and Frontiers, p228-264. Washington, D.C., ASM Press.
35
DePaola, A., C.A. Kaysner, J.C. Bowers, and D.W. Cook. 2000. Environmental investigations of Vibrio parahaemolyticus
in oysters following outbreaks in Washington, Texas, and New York (1997, 1998). Applied and Environmental
Microbiology, 66: 4649-4654.
36
Matsumoto, C., J. Okuda, M. Ishibashi, M. Iwanaga, P. Garg, T. Rammamurthy, H. Wong, A. DePaola, Y.B. Kim, M.J.
Albert, and M. Nishibuchi. 2000. Pandemic spread of an O3:K6 clone of Vibrio parahaemolyticus and emergence of related
strains evidenced by arbitrarily primed PCR and toxRS sequence analysis. Journal of Clinical Microbiology, 38: 578-585.
37
Kraa, E. 1995. Surveillance and epidemiology of food-borne illness in NSW, Australia. Food Australia, 47(9): 418- 423.
21
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
The determination of the dose-response relationship was based on the best available data.
Human volunteer studies were available for the construction of the dose-response curve for
V. parahaemolyticus, however, these studies characterize the dose-response relationship for
V. parahaemolyticus administered with a pH-neutralizing buffer rather than with a food matrix. The
data were analysed using curve-fitting routines to find a best fit for the Beta-Poisson dose-response
curve. Because of the limited amount of data available from human volunteer studies the resulting
dose-response relationship is uncertain. This uncertainty was accounted for by representing the doseresponse relationship in the form of a family of plausible data-derived dose-response curves
determined using resampling techniques. Figure 6.1 shows the most probable dose-response curve for
V. parahaemolyticus; however, the family of curves representing uncertainty that surrounds the curve
is not shown.
Probability of Gastroenteritis
0.1
0.01
0.001
0.0001
0.00001
0
10
12
22
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
The FDA-VPRA model was used as the base to accommodate data inputs from other countries. This
model incorporates all phases in the harvest - post-harvest consumption continuum in three modules
(Figures 6.2-6.4).
Figure 6.2 shows a conceptual model for the harvest module. Water temperature is the driving
input with regard to the initial numbers of V. parahaemolyticus in oysters. In the way the analysis is
constructed regional and seasonal temperature variations allow for a multi-year analysis that can
account for long-term temperature trends. Water salinity is shown in dotted lines to indicate that for
some model applications salinity may be another important input.
W ater salinity
total Vp/g
pathogenic V p/g
T im e to refrigeration
V p/g at harvest
A ir tem p erature
V p/g at 1st refrigeration
C oold ow n tim e
V p/g at cooldow n
S torage tim e
Figure 6.3 shows the conceptual model for post-harvesting practices. The post-harvest module
determines the role of post-harvest processing and handling on the numbers of pathogenic
23
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
V. parahaemolyticus at consumption. The bubble denoting "V.p/g at harvest" is the output of the
harvest model shown in Figure 6.2. Inputs on the time the oysters are out of the water and the air
temperature are used to predict growth of V. parahaemolyticus in the oysters. Growth continues as the
oysters are cooled but at a different rate. V. parahaemolyticus levels decrease during storage and the
storage time is therefore an input time that affects V. parahaemolyticus numbers.
Figure 6.4 represents the consumption module. The bubble denoting "path Vp/g (numbers) at
consumption" is the output of the post-harvest module. This number is multiplied by the number of
oysters per serving and the weight of the oysters to yield the ingested dose. This ingested dose is used
in the dose-response to calculate the risk of illness associated with the consumption of one oyster
meal.
V.p./g
(numbers) at
consumption
# oysters
per serving
Ingested
dose
Weight per
oyster (g)
RISK OF
ILLNESS
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
Japan
Based on the available data set41, the preliminary predictions of illness are shown in Table
6.2. The model predicted low levels of illness for November to April. The model was not run for the
months of May to October as oysters for raw consumption are not harvested during this period42.
It was difficult to compare this with epidemiological data for V. parahaemolyticus-related
oyster illnesses in Japan for a number of reasons. The Japanese surveillance system focuses mainly on
outbreaks of food-borne disease and therefore the number of laboratory confirmed reported illnesses
may not include sporadic cases or diffuse outbreaks and the extent of under-reporting is not known
(K Osaka, personal communication, 2002). In addition the food source of the illness may not always
be identified. However, in cases where oysters have been identified as the food source causing illness,
large variability in the annual number of V. parahaemolyticus-related oyster illnesses has been noted
over the last five years43. It is also worth noting that the model estimation is based on data (e.g. air and
water temperature, salinity) available from only one of the major harvesting areas and therefore does
not necessarily capture the situation in the different oyster growing areas in Japan.
TABLE 6.2: Preliminary predictions of V. parahaemolyticus illness in Japan associated with oyster
consumption
Number of
predicted illnesses
First quarter
(Jan-Mar)
4
Second quarter
(Apr-Jun)
1
(April only42)
Third quarter
(Jul-Sep)
042
Fourth quarter
(Oct-Dec)
19642
(Nov-Dec
only)
Total
201
Australia
Based on the available data set, the preliminary predictions of illness are shown in Table 6.3.
The model predicted more illnesses than the number of reported cases (J. Sumner, personal
communication, 2002). The application of United States of America surrogate data to a different
species of oyster, specifically the Sydney rock oyster, may have a role in the overestimation of risk.
TABLE 6.3: Preliminary predictions of V. parahaemolyticus illness in Australia associated with
oyster consumption
Number of
predicted illnesses
First quarter
(Jan-Mar)
157
Second quarter
(Apr-Jun)
28
Third quarter
(Jul-Sep)
10
Fourth quarter
(Oct-Dec)
33
Total
228
New Zealand
The model predicted more illnesses than the number of reported cases (D.J. McCoubrey,
personal communication, 2002) (Table 6.4). As extensive use of surrogate data from the United States
41
25
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
of America was necessary as inputs for some of the parameter required to run the model, the true risk
may be much lower than that predicted.
TABLE 6.4: Preliminary predictions of V. parahaemolyticus illness in New Zealand associated with
oyster consumption
Number of
predicted illnesses
First quarter
(Jan-Mar)
13
Second quarter
(Apr-Jun)
17
Third quarter
(Jul-Sep)
0
Fourth quarter
(Oct-Dec)
5
Total
35
Canada
The preliminary results (Table 6.5) indicate that model predicted cases of illness that are
relatively close to the number of reported cases 4445). The proximity of the Canadian harvesting waters
to one of the regions of the United States of America that was modelled allows greater confidence in
these predictions. It should be noted that the model did not consider the mitigation to cool oysters
immediately after harvest that was introduced in the Canadian oyster industry in 2000, as the data
used was collected prior to the implementation of this measure.
TABLE 6.5: Preliminary predictions of V. parahaemolyticus illness in Canada associated with oyster
consumption
Number of
predicted illnesses
First quarter
(Jan-Mar)
0
Second quarter
(Apr-Jun)
1
Third quarter
(Jul-Sep)
7
Fourth quarter
(Oct-Dec)
0
Total
8
Number of
predicted illnesses
First quarter
(Jan-Mar)
40
Second quarter
(Apr-Jun)
1587
44
Third quarter
(Jul-Sep)
3881
Fourth quarter
(Oct-Dec)
376
Total
5884
Cato, J.C. 1998. Economic values associated with seafood safety and implementation of seafood, Hazard Analysis Critical
Control Point (HACCP) programmes. FAO Fisheries Technical Paper. No. 381. Rome, FAO. 1998.
45
Anonymous, 1997, Canada Communicable Disease Report - Volume 23-19, October 1, 1997.
46
Mead, P.S., Slutsker, L., Dietz, V., McCaig, L.F., Bresee, J.S., Shapiro, C., Griffin, P.M. and Tauxe R.V. 1999. FoodRelated Illness and Death in the United States Emerging Infectious Diseases, 5 (5), 607-625.
26
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
47
Mead, P.S., Slutsker, L., Dietz, V., McCaig, L.F., Bresee, J.S., Shapiro, C., Griffin, P.M. and Tauxe R.V. 1999. FoodRelated Illness and Death in the United States Emerging Infectious Diseases, 5 (5), 607-625
27
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
Methodology for estimating the ratio of reported to total cases of illness. Appropriate
methodology needs to be officially developed and applied in countries that wish to compare
the number of illnesses predicted by risk assessment to the number of reported & recorded
cases of illness.
Data sources that can indicate whether or not the model is succeeding. Validation of the risk
assessment model should be attempted at as many intermediate stages as is practical.
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
the harvest site. Following initial sampling (Harvest stage), the remaining clams were transported to
the local market area, which was located close to the laboratory. A sample of clams were examined at
this point to represent the Retail stage. Thereafter, the clams were maintained outside of the
laboratory for a period of time to simulate the transportation step; these were subsequently examined
in the laboratory. Typically, the clams are cooked in the home by boiling briefly (in some cases with
insufficient heating). The Cook (Boiling) stage was simulated in the laboratory and the clams were
tested thereafter. To obtain consumption data, local people were interviewed on the frequency and
quantity of bloody clams consumed.
TABLE 6.6: Results of the study on isolation of V. parahaemolyticus from seafood and the most
common strain profiles of isolates from clinical specimens and seafood
Isolation of pathogenic strains of
V. parahaemolyticus
O1:K25 tdh+,
trh-
13/268 (4.4%)
0/50
0/9
0/100
8(62%)
0
0
0-
2(15%)
0
00-
192 (65%)
22(7.5%)
Seafood samples*
Shellfish(bivalves)
Shrimp
Crab
Fish
Clinical samples**
*Samples were examined over a four year period from 1998 to 2001. During the first year of the study period pathogenic
V. parahaemolyticus were only isolated from shellfish. Therefore, during the subsequent years of the survey, efforts focussed
mainly on detection of pathogenic V. parahaemolyticus in shellfish samples.
** V. parahaemolyticus was isolated from 317 diarrhoea specimens out of a total of 11 375 samples that were examined
during a survey sporadic cases of illness with diarrhoea in 1999. Specimens came from different patients in two big hospitals
in the province. Of the 317 cases confirmed positive for V. parahaemolyticus, 294 of these were confirmed to be pathogenic
strains of V. parahaemolyticus.
Harvest
Temperature
Prevalence
Concentration
transportation
Time
Temperature
Retail
Temperature
Prevalence
Concentration
Cooking
Temperature
Prevalence
Concentration
Quantity
Frequency
transportation
Time
Temperature
Consumption
FIGURE 6.5: Schematic representation of the exposure model developed for the risk assessment of
V. parahaemolyticus in bloody clams.
29
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
30
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
The results are restricted to a single food item, and the sample size may not be sufficiently
large. Therefore, the data presented in table 6.6 should be interpreted with caution. Furthermore, the
study on survival of V. parahaemolyticus from harvest to consumption was carried out for only a three
month period in one specific area in Thailand. More data are needed for other months and other areas.
Because the cooking (by boiling) module was developed based on experimental data using
fixed time / temperature values within a very limited range, scenario analysis with different time /
temperature combinations are impossible. Also the consumption survey was carried out on a small
group of people working within the same environment and therefore may not be representative of the
region as a whole.
The cross-contamination model was not applied in this risk-assessment, because of lack of
data and appropriate models for cross-contamination. Due to insufficient epidemiological data, model
validation could not be undertaken.
6.1.4.9 Gaps in the data
48
WHO. 2001. Joint FAO/WHO Expert Consultation on Risk Assessment of Microbiological Hazards in Foods: Hazard
identification, exposure assessment and hazard characterization of Campylobacter spp. in broiler chickens and Vibrio spp. in
seafood, WHO Headquarters, Geneva, Switzerland 23 - 27 July 2001. WHO 2001.
http://www.fao.org/es/ESN/food/risk_mra_campylobacter_en.stm / http://www.who.int/fsf/Micro/index.htm
31
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
6.1.5.2 Scope
This work focused on describing the
V. parahaemolyticus from harvest to consumption.
possible
contamination
of
finfish
by
32
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
49
Mead, P.S., Slutsker, L., Dietz, V., McCaig, L.F., Bresee, J.S., Shapiro, C., Griffin, P.M. and Tauxe R.V. 1999. FoodRelated Illness and Death in the United States Emerging Infectious Diseases, 5 (5), 607-625
50
Dalsgaard, A., Hoei, L., Linkous, D. and Oliver, J.D. 2001. Vibrio vulnificus. In Y.H. Hui, M.D. Person and J. R. Gorham
(eds). Foodborne disease handbook, vol 1 Bacterial Pathogens, New York, Marcel Dekker Inc.
51
Oliver, J. D., and Kaper, J.B. 1997. Vibrio Species. In M. P. Doyle, L. R. Beuchat, and T. J. Montville, eds. Food
Microbiology: Fundamentals and Frontiers, p228-264. Washington, D.C., ASM Press.
33
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
34
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
total Vv/g
POS T-HARVES T
Vv/g at harvest
Time to refrigeration
Air temperature
Vv/g at 1st refrigeration
Cooldown time
Vv/g at cooldown
S torage time
Vv/g (numbers) at consumption
S usceptible population
FIGURE 6.6: Schematic diagram of the V. vulnificus conceptual risk assessment model showing
integration of all the modules.
35
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
is quite flexible and most model inputs could be easily adapted to fit specific situations if appropriate
data were available.
The dose-response approach used in this assessment was a curve-fitting of the data to a betapoisson model. While the beta-poisson model was selected any other empirical model that fits the data
could be used. Extrapolation of the beta-poisson parameters used in this analysis beyond the range of
normal consumption would be inappropriate.
The application of this model to predict the risk of V. vulnificus illnesses from seafoods other
than molluscan shellfish was limited as the ecology of this bacterium differs considerably as do
industry practices and consumer handling. However, the dose-response relationship could be useful in
determining the risk of other seafoods if V. vulnificus levels in these products were known at the point
consumption. The accuracy of these assessments would depend on the extent of matrix effects on the
dose-response.
The model does not account for variation of strain virulence. Regional or seasonal variation in
V. vulnificus virulence could alter the current dose-response and affect illness estimates.
This assessment was based on the distribution of at risk individuals52 in the population of the
United States of America and this parameter would need to be redefined on a country by country basis
depending on the size and characterization of the at risk population.
Spring
Summer
Fall
Season
FIGURE 6.7: Predicted and observed numbers of V. vulnificus per gram oyster on a seasonal basis.
Since the dose-response data was generated in part using monthly illness rates in the United
States of America, this data, in its current format, cannot be used to validate the model. However,
illness data from that country may be useful for validation of the risk characterization in a different
format (i.e. retrospective analysis of annual illness rates before and after specific mitigations). In 1997
the Interstate Seafood Sanitation Conference (ISSC) in the United States of America adopted a time52
Individuals with predisposing conditions which include insulin-dependant diabetes, liver disease (cirrhosis), gastric
acidity, cancer, hepatitis B & C, kidney disease, haemochromatosis, AIDS, being immunocompromised due to
treatment/surgery, asthma, rheumatoid arthritis, psoriatic arthritis, lupus, polymylagia rheumatica, giant cell arthritis and
being a transplant recipient.
36
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
temperature matrix for reducing the time to first refrigeration of oysters from 20 to 10h under certain
circumstances. The model could be used to analyse the effect on exposure and predicted illness and
these could be compared to illness rates before and after adoption of the time-temperature matrix.
6.1.6.9 Gaps in the data
In the course of this work the following data gaps were identified.
There was sufficient exposure data available for modelling the risk of V. vulnificus illness
from consumption of raw Gulf Coast oysters in the United States of America using the
proposed approach but this data especially numbers of V. vulnificus at harvest is also lacking
in most other countries.
There is a lack of reliable markers for virulence determination of V. vulnificus, thus requiring
the assumption that all strains are virulent.
The incidence of specific risk factors in the population consuming a seafood of interest and
exposure associated with this seafood are the primary data needed for applying this model to
other countries.
Validation of the model in a given region or country would require epidemiological data on
the incidence of primary septicaemia caused by V. vulnificus on a monthly basis.
FAO (1999) World production of fish, crustaceans and molluscs reached 126.2 million tonnes in 1999, an increase of 7.2
% above the 1998 level. http://www.fao.org/fi/trends/worldprod99e.asp .
54
Desmarchelier, P.M. 1997. Pathogenic Vibrios. In A.D. Hocking, G. Arnold, I. Jenson, K. Newton and P. Sutherland, eds.
Foodborne Microorganisms of Public Health Significance 5th Edition, p 285 -312. North Sydney, Australian Institute of
Food Science and Technology Inc.
37
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
Outbreaks of cholera have been associated with consumption of seafood including oysters, crabs and
shrimp (Oliver and Kaper, 199755). The largest outbreak was a pandemic in South America in the
early 1990s when V. cholerae O1 caused more than 400,000 cases and 4,000 deaths, in Peru (Wolfe,
199256). Contaminated water used to prepare food, including the popular, lightly marinated fish
ceviche, was associated with the outbreak.
Cholera occurs in areas with inadequate sanitary conditions and infrastructure and is
associated with faecal contamination of water and foods. V. cholerae is widely distributed in coastal
and estuarine environments all over the world and there exists over 170 serotypes of V. cholerae.
According to WHO definitions57, only serotypes O1 and O139 are the causes of cholera.
Ability to produce cholera toxin (CT) is the determining virulence factor for causing cholera.
However, environmental strains of V. cholerae O1 have often been shown to be non-toxigenic.
Though some strains of non-O1/O139 V. cholerae may also cause gastroenteritis, the disease is of a
mild to moderate severity58. Choleragenic V. cholerae is susceptible to inactivation by cooking. Most
of the risk associated with choleragenic V. cholerae comes from the consumption of raw seafood or
from cross-contamination of the foods by food handlers or contaminated water.
Accordingly, this risk assessment considers only choleragenic V. cholerae O1 and O139.
6.1.7.4 Hazard characterization
V. cholerae O1 and O139 have both pathogenic and non-pathogenic forms based on the
presence of specific virulence genes, ctx (cholera toxin gene). Infection by choleragenic V. cholerae
O1 and O139 is characterized by an acute gastroenteritis. Therefore, the end-point of the doseresponse curve was defined as gastroenteritis.
Human volunteer studies were available for the construction of the dose-response curves for
V. cholerae O1. Reasonable Beta-Poisson dose-response parameters were obtained from data sets;
however, the human volunteer studies characterize dose-response relationships for pathogens
administered with a pH-neutralizing buffer rather than for pathogens administered with a food matrix.
In normochlorohydric adult volunteers, doses of up to 10 11 choleragenic V. cholerae given without
buffer or food did not reliably cause illness, whereas doses of 10 4 to 10 8 organisms given with
NaHCO3 (sodium bicarbonate) resulted in diarrhoea in 90% of individuals. Dose-response curves
(Figure 6.8) show that a high dose of V. cholerae O1 (10 6) was normally needed to cause illness
when V. cholerae are consumed in food. In populations not exposed to choleragenic V. cholerae all
age groups are equally susceptible. Immunity seems to be serotype specific.
6.1.7.5 Exposure assessment
This risk assessment includes aquacultured and wild-caught warm-water shrimp.
Choleragenic V. cholerae O1 and O139 generally occur in waters with salinity's between 0.2 to 20
ppt. Therefore, water and shrimp from offshore waters have not been found to contain choleragenic
V. cholerae. Thus, it is assumed that any presence of choleragenic V. cholerae in offshore wild caught
shrimp is caused by post-harvest cross-contamination. Though the presence of choleragenic
V. cholerae in aquaculture environments is very rare, the model assumes that such choleragenic
V. cholerae strains could be present in shrimp at levels similar to those found in coastal waters of
cholera endemic countries.
The model developed was based on shrimp handling, processing and storage practices in units
approved for export of shrimp (Figure 6.9). Such approval is based on sanitary requirements as
described in Good Manufacturing Practices (GMP) and Good Hygienic Practices (GHP). Shrimp
55
Oliver, J. D., and Kaper, J.B. 1997. Vibrio Species. In M. P. Doyle, L. R. Beuchat, and T. J. Montville, eds. Food
Microbiology: Fundamentals and Frontiers, p228-264. Washington, D.C., ASM Press.
56
Wolfe, M. 1992. The effects of cholera on the importation of foods: Peru- a case study. PHLS Microbiology Digest, 9: 4244.
57
WHO Fact Sheet N107
58
Morris, J.G. Jn. 1990. Non-O group 1 Vibrio cholerae: a look at the epidemiology of an occasional pathogen.
Epidemiological Review, 12: 179-191.
38
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
intended for export are generally iced immediately after harvest and transported in ice to certified
processing units that meet GHP/GMP requirements. However, a worst-case scenario of shrimp being
processed in non-approved units was also considered.
The major factors influencing the numbers of choleragenic V. cholerae in shrimp are time and
temperature during handling, processing and storage. In the absence of available data it was necessary
to make assumptions on distributions of time and temperature under such conditions. Adequate data
were available on the effect of washing, freezing and cooking on the numbers of choleragenic
V. cholerae in shrimp. In particular, the duration of frozen storage before consumption will cause a
significant reduction in numbers of choleragenic V. cholerae. Limited information was available on
the levels of faecal cross-contamination during handling and multiplication of choleragenic
V. cholerae in raw shrimp. The model takes into account the pronounced reduction in the levels of
choleragenic V. cholerae that would occur during cooking of shrimp either before export or before
consumption. It also assesses the risk if shrimp were consumed raw or inadequately cooked in the
importing country.
1.00
probability of illness
0.80
Classical with Food Matrix
(Cash)
El Tor (Levine 1988)
0.60
0.40
0.20
0.00
1.E+00
1.E+02
1.E+04
1.E+06
1.E+08
1.E+10
1.E+12
cholera dose
39
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
V. cholerae / cm on
fingers of handlers
V. cholerae / g in ice
V. cholerae / ml in
water
Shrimp washed,
peeled, graded,
packed and frozen
V. cholerae / g
No. of V. cholerae
ingested
FIGURE 6.9: Conceptual model for risk assessment of choleragenic V. cholerae in warm water
shrimp for export.
40
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
The following data gaps were identified in the course of the work.
Data on the levels of choleragenic V. cholerae O1 and O139 in natural waters and aquaculture
environments.
Data on the multiplication of choleragenic V. cholerae in cooked and raw shrimp.
Data on levels of faecal cross contamination during handling of shrimp.
Data to clarify the dose-response when the El Tor form is ingested in food.
41
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
59
WHO. 2001. Report of the Joint FAO/WHO Expert Consultation on Risk Assessment of Microbiological Hazards in
Foods; Hazard identification, exposure assessment and hazard characterization of Campylobacter spp. in broiler chickens
and Vibrio spp. in seafood. WHO Headquarters, Geneva, Switzerland 23 - 27 July 2001. WHO 2001.
http://www.fao.org/es/ESN/food/risk_mra_campylobacter_en.stm / http://www.who.int/fsf/Micro/index.htm
42
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
43
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
make the software models available together with guidance on their use. They should also provide
technical assistance to developing countries wishing to extend the assessments to local needs.
These risk assessments could be used to support relevant risk management decisions. The
output from the assessments should also be used to formulate and target research needs, e.g. to fill
identified data gaps.
6.4
44
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
estimates could also be obtained of the proportion of harvest lost by application of a particular
scenario.
Some of the listed mitigations are also used in combination, e.g. hydrostatic pressure and
freezing; depuration and hydrostatic pressure or pasteurization.
TABLE 6.7: The comparative effectiveness of a number of mitigation strategies in reducing Vibrio
spp..
Mitigation
Hydrostatic pressure
Rapid cooling
Irradiation
Pasteurization
Freezing and thawing
Depuration
Relay at high salinity for 2 weeks
(for V. vulnificus)
Commercial heat-treatment
+
++
+++
no effect
some reduction
moderate reduction
significant reduction
45
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
Richards, G. P. 1988. Microbial purification of shellfish: A review of depuration and relaying. J. Food
Protect. 51:218-251.
Son, N. T., and G. H. Fleet. 1980. Behaviour of pathogenic bacteria in the oyster, Crassostrea
commercialis, during depuration, re-laying, and storage. Applied and Environmental
Microbiology, 40:994-1002.
Question 3
For Vibrio parahaemolyticus - Are food-borne illnesses caused by the heat
resistant toxin produced by the pathogen or by the pathogen itself?
Response from the consultation: The illness is caused by the toxin but only if this is
produced in the intestine following colonization by a strain producing TDH, TRH or both toxins.
Question 4: What is the availability of methods of analysis for Vibrio parahaemolyticus
toxin gene (tdh)?
Response from the consultation: Both tdh and trh genes can be detected using PCR with
relevant primers and by membrane filtration-hybridization methods with non-isotopic oligonucleotide
or PCR-generated probes. For quantification, PCR methods can be applied in an MPN format whereas
membrane filter-hybridization can be used for direct colony enumeration. PCR and colony
hybridization procedures are also available for the thermolabile haemolysin gene (tlh) for determining
V. parahaemolyticus species. As with conventional methods, there is scope for standardization and/or
the determination of the relative performance of current methods.
WHO 2001. Report of the Joint FAO/WHO Expert Consultation on Risk Assessment of Microbiological Hazards in
Foods; Hazard identification, exposure assessment and hazard characterization of Campylobacter spp. in broiler chickens
and Vibrio spp. in seafood. WHO Headquarters, Geneva, Switzerland 23 - 27 July 2001. WHO 2001.
http://www.fao.org/es/ESN/food/risk_mra_campylobacter_en.stm / http://www.who.int/fsf/Micro/index.htm
46
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
7 Conclusions
The assessments presented at this expert consultation were in varying degrees of completion.
The risk assessments for Campylobacter spp. in broiler chickens, V. vulnificus and
V. parahaemolyticus in oysters, are most suited at the present time for use by risk managers to help
them make informed risk management decisions. The models use a modular approach which lends
flexibility for their use in assessing production systems not specifically covered in the current models.
The utilization of the models was demonstrated by a number of examples showing how certain
mitigation strategies may lead to a reduction in the number of cases of disease. The models are still
evolving and once fully completed will become a useful tool for risk managers.
Since the last consultation, the hazard characterization and exposure assessment were merged
to produce a preliminary risk characterization. The risk characterization can provide risk managers
with the ability to gain insight into the relative effectiveness of various mitigation strategies. Future
activities will focus on ensuring the full potential of the risk assessment is achieved. In addition, a full
exploration of the model will be undertaken to fully understand the scope and limitations
8 Recommendations
47
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
provide assistance to developing countries to gather quantitative information for risk assessment.
Appropriate resources should be provided to developing countries to train and undertake risk
assessments and those wishing to extend the risk assessments to local needs.
encourage microbiologists to develop and employ techniques to differentiate pathogenic and nonpathogenic strains.
encourage governments that have set or intend to set targets for food-borne diseases addressed in
this report to use the models presented to inform their decisions in setting appropriate food safety
objectives and intervention strategies.
commission a document to assist a wider readership in understanding this report. The
recommended document would describe for non-specialists appropriate information on;
i) shellfish and poultry e.g. production methods, processing, economics and legislation.
ii) Vibrio spp. and Campylobacter spp. (surveillance, epidemiology and microbiology).
iii) risk assessment (its approach, components and how it can answer risk managers
questions).
facilitate hands-on demonstrations for identifying and collecting relevant data for risk assessment
for specific commodity/pathogen combinations including risk characterization using various
modelling software.
develop new ways of communicating the concepts of risk assessments to users.
promote utilization of the valuable framework models that have now been established to
undertake further risk assessments. It would be relatively easy to use modules from these risk
assessments to develop a risk assessment of Salmonella in shrimps. The experts consider this
would be a valuable risk assessment for future consideration.
consider, for future risk assessment activities, the involvement of scientific experts to serve as a
standing advisory resource group to the risk assessment drafting group, for the duration of a
specific work assignment. This would be a significant complementary activity to the more formal
in-person experts meetings and would certainly take greater advantage of the intellectual
resources of the expert community through their knowledge of and access to relevant data,
references and network of colleagues.
encourage the submission of appropriate aspects of the work to peer-reviewed technical
publications and the seafood and poultry trade press.
make the software models available in a format usable by other professionals and provide
appropriate guidance on their use.
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Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
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Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
50
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
Title
Authors
(in alphabetical order)
MRA 02/01
MRA 02/02
51