Available online at www.sciencedirect.

com

International Journal for Parasitology 38 (2008) 1359–1370

www.elsevier.com/locate/ijpara

Invited Review

Control of the risk of human toxoplasmosis transmitted by meat

Aize Kijlstra a,b,*, Erik Jongert c
a
Animal Sciences Group, Wageningen UR, P.O. Box 65, 8200 AB Lelystad, The Netherlands
b
Department Ophthalmology, Maastricht University, Maastricht, The Netherlands
c
Laboratory for Toxoplasmosis, Pasteur Institute of Brussels, Scientific Institute of Public Health, Belgium, Engelandstraat 642, 1180 Brussels, Belgium

Received 14 May 2008; received in revised form 25 June 2008; accepted 29 June 2008

Abstract

One-third of the human world population is infected with the protozoan parasite Toxoplasma gondii. Recent calculations of the dis-
ease burden of toxoplasmosis rank this foodborne disease at the same level as salmonellosis or campylobacteriosis. The high disease bur-
den in combination with disappointing results of the currently available treatment options have led to a plea for more effective
prevention. In this review we describe Toxoplasma as a hazard associated with the consumption of undercooked meat or meat products
and provide an analysis of the various options to control the risk of human toxoplasmosis via this source. Monitoring and surveillance
programs may be implemented for pre-harvest control of Toxoplasma infection of farm animals, with the reduction of environmental
oocyst load as the most important milestone. Alternatively, Toxoplasma safe meat can be obtained through simple post-harvest decon-
tamination procedures, whereby freezing the meat may currently be the best option, although new technologies using irradiation or high-
pressure treatment may offer promising alternatives. Influence of culture, religion and food handling customs may predispose a certain
type of meat as an important source of infection, indicating that prevention needs to be tailored according to social habits in different
regions in the world. The rationale for more stringent control measures to prevent toxoplasmosis both from disease and economic points
of view is emphasized.
Ó 2008 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

Keywords: Toxoplasma gondii; Meat; Parasite inactivation; Food safety

1. Introduction ences throughout the world, and have extensively dealt

Toxoplasmosis is a disease caused by the protozoan par-
asite Toxoplasma gondii (Montoya and Liesenfeld, 2004)
and infection with the parasite is ubiquitous throughout
the world. Less than 20% of individuals develop overt dis-
ease including fever, lymph node enlargement or intraocu-
lar inflammation (Holland, 2003; AFSSA, 2005). Details
concerning both asymptomatic and symptomatic infection
and the risks of transmission from animals to humans in
the USA were recently reviewed by Dubey and Jones
(2008). In the review presented here we have focussed on
meat as a source of infection, highlighting regional differ-
*
Corresponding author. Address: Animal Sciences Group, Wageningen
UR, P.O. Box 65, 8200 AB Lelystad, The Netherlands. Tel.: +31 320
238095; fax: +31 320238050.
E-mail address: aize.kijlstra@wur.nl (A. Kijlstra).
with pre- and post-harvest mechanisms of control. Further-
more, the rationale for more stringent control measures to
prevent toxoplasmosis both from disease and economic
points of view is emphasized.
More emphasis on prevention is needed because cur-
rently available antibiotic therapy seems to have little effect
on mother-to-child transmission, and whether treatment
affects the clinical manifestations in the newborn with con-
genital toxoplasmosis (CT) is still under debate (Thibaut et
al., 2007). The effectiveness of antiparasitic treatment for
chronic ocular toxoplasmosis has also not yet been for-
mally demonstrated (Rothova et al., 1993; Stanford et
al., 2003). Treatment failure may be related to late timing
of treatment and the fact that currently available drugs
cannot reach the parasite stage within tissue cysts (Dubey,
1996; Gormley et al., 1998). In humans, primary infection
with the parasite before pregnancy provides complete

0020-7519/$34.00 Ó 2008 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.ijpara.2008.06.002
1360 A. Kijlstra, E. Jongert / International Journal for Parasitology 38 (2008) 1359–1370
protection against congenital toxoplasmosis (Montoya and
Liesenfeld, 2004).
Despite the low incidence, the economic impact of CT is
high due to severity of infection, associated complications,
treatment and social costs. Costs of CT were estimated up
to US$ 1.26 million per case, and were mainly attributed to
medical costs, annual productivity losses, special education
and residential care costs (Roberts et al., 1994). The total
economic impact of CT in the USA has been estimated
to be as high as US$ 7.7 billion per year, which makes it
the second most important foodborne infection for humans
after salmonellosis (Buzby and Roberts, 1996). In the UK,
the annual economic impact has been estimated at US$ 12
million (Roberts and Frenkel, 1990). The costs associated
with acquired toxoplasmosis have never been addressed,
despite the fact that up to 20% of infected individuals
may develop clinical complications, probably leading to
one or more days of work missed in mild cases (AFSSA,
2005). Infectious mononucleosis such as illness due to T.
gondii is often missed as a diagnosis and it may therefore
remain an under-reported disease entity (Bottieau et al.,
2006).
Despite the fact that disease burden of toxoplasmosis is
comparable to that of other foodborne diseases such as sal-
monellosis or campylobacteriosis, toxoplasmosis has
received little attention from policy makers in past years
(Mead et al., 1999; Vaillant et al., 2005; Kemmeren et al.,
2006; Havelaar et al., 2007). Ocular toxoplasmosis was tra-
ditionally considered to be a manifestation of congenital
disease. Recent insight, however, shows that most cases
are due to acquired disease which means that not only
pregnant women, but the general population, are at risk
(Glasner et al., 1992; Gilbert and Stanford, 2000). In the
UK, 50% of children presenting with chorioretinitis are
due to Toxoplasma infection after birth (Stanford et al.,
2006). Holland has estimated that approximately 2% of
infected individuals will have ocular involvement (Holland,
2003).
Sources of human infection with T. gondii are oocysts
shed in faeces of infected felines and tissue cysts from
infected meat animals. Transmission of oocysts occurs by
water and soil, and thus might also be transferred via
raw vegetables and fruit. Inhalation of dust containing
oocysts has also been implicated as a means of transmis-
sion (Teutsch et al., 1979).
2. Indications that meat is a main cause of T. gondii infection
in humans
The source by which an individual has become infected
with T. gondii cannot be discriminated by diagnostic tests.
In Europe, three large case-control studies have pinpointed
uncooked meat as the most important risk factor for preg-
nant women (Kapperud et al., 1996; Baril et al., 1999;
Cook et al., 2000) (Table 1). The largest study estimated
that consumption of not-well-cooked meat was the cause
of infection in 30–60% of pregnant women with acute toxo-
Table 1
The risk of Toxoplasma gondii infection in association with the consump-
tion of undercooked or raw meat from various species
Type of meat Kapperud Baril Cook
et al. (1996) et al. (1999) et al. (2000)
Mutton/Lamb 4.1 (2.1–63.1) 3.1 (0.85–14) 3.1 (1.4–7.2)
Pork 3.4 (1.1–10.4) No increased risk
Beef No increased risk 5.5 (1.1–27) 1.73 (1.1–7.2)
Poultry No increased risk
Othera 4.12 (1.6–10.9)
Number of cases 63 80 252
Number of controls 128 80 858
Multivariate analysis data are represented from three large case-control
studies and depicted as odds ratio plus confidence intervals (in brackets).
a
Venison, horse, rabbit, whale, game birds.
plasmosis (Cook et al., 2000). Small outbreaks of toxoplas-
mosis have been associated with the consumption of raw
meat in Korea, USA France, French Guiana and
New Zealand (Choi et al., 1997; Ross et al., 2001; Carme
et al., 2002; Lake et al., 2002). However, the fact that strict
vegetarians become infected with T. gondii shows that
oocyst contamination still plays an important role in infec-
tion (Hall et al., 1999; Roghmann et al., 1999).
The observed decline in Toxoplasma seroprevalence as
noted in many developed countries over past decades has
been attributed to the introduction of modern farming sys-
tems resulting in a lower prevalence of Toxoplasma cysts in
meat in combination with an increased use of frozen meat
by consumers (Tenter et al., 2000; Kortbeek et al., 2004;
AFSSA, 2005; Diza et al., 2005; Jones et al., 2007). A
few studies have been able to link T. gondii outbreaks to
oocyst contamination of drinking water in the United
States, Canada and Brazil (Bowie et al., 1997; Bahia-
Oliveira et al., 2003; Dubey, 2004). However, a recent study
has shown that T. gondii oocyst contamination of the envi-
ronment is mostly restricted to cat defecation sites (Afonso
et al., 2008). Less than 0.9% of cats have been found to
actively shed Toxoplasma-like oocysts, (Childs and Seegar,
1986; Svobodova et al., 1998; Dabritz et al., 2007; Schares
et al., 2008), and this may be an over-estimation since T.
gondii oocysts are difficult to distinguish from Hammondia
hammondi oocysts by microscopical analysis (Schares et al.,
2008).
Although it is generally assumed that raw vegetables are
a source of contamination with T. gondii in humans, to
date no experimental data or studies are available to sup-
port this route of infection. One study has shown that ber-
ries experimentally spiked with oocysts can pass T. gondii
infection to mice (Kniel et al., 2002).
3. Food animal species involved in T. gondii transfer to
humans
Experimental infections of food animals such as cattle,
pigs, sheep and goats, have shown that these animals are
susceptible to T. gondii contamination by intake of oocysts
 A. Kijlstra, E. Jongert / International Journal for Parasitology 38 (2008) 1359–1370
or tissue cysts, and that following experimental infection T.
gondii can be isolated from their tissues, with the exception
of beef (Dubey et al., 1980, 1984; Blewett et al., 1982;
Dubey, 1983, 1986a, 1988; McColgan et al., 1988; Lunden
and Uggla, 1992; Dubey and Thulliez, 1993; Esteban-
Redondo et al., 1999).
In response to natural infection, most farm animals that
are seropositive for T. gondii have been shown to harbour
infectious parasites in their meat, with the exception of beef
(Tenter et al., 2000; Zia-Ali et al., 2007). Undercooked
meat has thus been considered a main source of infection.
A comprehensive review on T. gondii induced pathology in
livestock animals and their role in the transmission of T.
gondii to humans in the USA was recently reviewed by
Dubey and Jones (2008).
3.1. Sheep
All case-control studies have identified the consumption
of mutton/lamb meat as a highly significant risk factor for
contracting T. gondii infection in pregnant women.
Depending on the environmental load with oocysts, sheep
can readily acquire toxoplasmosis (Skjerve et al., 1998).
Seroprevalence of T. gondii in sheep can be as high as
92% in certain European countries (Tenter et al., 2000).
Raw or undercooked lamb meat is considered a delicacy
in certain countries such as France and is therefore consid-
ered an important source of infection in that country
(AFSSA, 2005). Adult sheep meat is often well cooked
and therefore probably poses a smaller risk of infection
to the consumer than lamb meat. Recently a large-scale
screening of sheep farms has shown that 3.4% of sheep
were shedding T. gondii in their milk (Fusco et al., 2007).
3.2. Pigs
Outbreaks of acute toxoplasmosis have been described
following the consumption of uncooked pork (Choi et
al., 1997). Due to major changes in animal production
hygiene, the rate of T. gondii infection of pork meat has
dropped dramatically (Tenter et al., 2000; Dubey and
Jones, 2008). Currently, modern production systems have
virtually eliminated T. gondii infection in pigs (Vanknapen
et al., 1995; Davies et al., 1998; Kijlstra et al., 2004a; van
der Giessen et al., 2007). Pork meat is therefore not the
main source of infection any more in many parts of the
world. However, increasingly popular animal friendly pro-
duction systems with increased risk of exposure to T. gondii
may cause a re-emergence of pork meat as an infectious
meat source (Kijlstra et al., 2004a; Schulzig and Fehlhaber,
2006; van der Giessen et al., 2007).
3.3. Goats
Seroprevalence for T. gondii in goats can be as high as 77%
and is dependent on the presence of oocysts in the environ-
ment, housing and climatic conditions (Tenter et al., 2000).
1361
Although studies have reported the isolation of T. gondii
from caprine tissues, no large-scale prevalence data are avail-
able on the presence of parasites in goat meat products
(Dubey, 1980, 1981; Sharma et al., 2003). Small ruminants
such as goats are an important source of meat and milk in
many undeveloped countries and may play a role as a source
of infection for humans residing in these areas (Shrestha and
Fahmy, 2005). Consumption of raw goat’s milk and milk
products has been linked to cases of toxoplasmosis in
humans and pigs (Riemann et al., 1975; Sacks et al., 1982;
Skinner et al., 1990; Meerburg et al., 2006).
3.4. Beef
Although epidemiological studies have shown that the
consumption of raw or undercooked beef is considered a
risk for T. gondii infection in humans (Baril et al., 1999;
Cook et al., 2000), and that seroprevalence can be high
in bovine (up to 92% has been reported) (Vanknapen et
al., 1995; Tenter et al., 2000; Sroka, 2001; More et al.,
2008), conclusive evidence is lacking to correlate this with
the actual presence of infectious parasites in edible tissues
from naturally infected beef (Dubey, 1986b). In a study
by Hellman and Tauscher the presence of viable parasites
in 170 commercially obtained beef cuts was evaluated but
none tested positive (Hellmann and Tauscher, 1967).
Drinking unpasteurized (cow) milk was not associated with
T. gondii infections (Kapperud et al., 1996). Outbreaks
have been reported following consumption of raw beef,
although doubt was raised whether the meat was unadul-
terated (Dubey and Jones, 2008).
3.5. Poultry
Seroprevalence of up to 65% in free ranging chickens
has been reported and the presence of the parasite in meat
could be shown in 81% of seropositive animals (da Silva et
al., 2003; Lehmann et al., 2006). Free ranging chickens,
especially in developing countries, may be considered as
an important source of T. gondii infection in humans. In
the Western world, commercially produced free ranging
chickens intended for meat consumption (broilers) have a
limited life span and to date no recent data are available
concerning T. gondii seroprevalence in these chickens, how-
ever it can be expected that poultry kept outside has a
higher chance of being infected with the parasite (Dubey
et al., 2004). Chicken meat is mostly well cooked for
consumption.
3.6. Horses
Horses can become infected with T. gondii and in certain
regions of the world, up to 90% of the animals were shown
to be seropositive (Tassi, P., 2006. Toxoplasma gondii infec-
tion in horses- a serological survey in horses slaughtered
for human consumption in Italy. In, Toxo & Food 2006,
Palermo, Italy, pp. 96–97; Tassi, 2007). Presence of cysts
1362 A. Kijlstra, E. Jongert / International Journal for Parasitology 38 (2008) 1359–1370
has been shown in edible tissues from horses (Alkhalidi and
Dubey, 1979). The role of horses as a source of T. gondii
infection depends on regional preferences for horse meat,
the preparation method and the seroprevalence of horses
used for consumption (Gill, 2005; Tassi, 2006). Raw or
undercooked horse meat is frequently consumed in coun-
tries such as Belgium, Italy, France and Japan (Gill, 2005).
3.7. Game
An emerging risk is the increasing popularity of game
meat such as roe deer, wild boar or kangaroo, and a study
from the European Food Safety Authority has recently
estimated that approximately half of the game produced
in Europe may be seropositive for T. gondii (EFSA,
2007). Epidemiological studies and several outbreaks have
identified the handling and consumption of raw or under-
cooked game as a source of toxoplasmosis (McDonald et
al., 1990; Cook et al., 2000; Ross et al., 2001; Carme et
al., 2002; Dubey and Jones, 2008). Due to high demand
in certain countries there is an intense trade of both farmed
and natural game, partly via frozen import or fresh meat
imports (Reinken, 1998). Serological studies showed that
between 13% and 39.2% of European roe deer (Capreolus
capreolus) were seropositive for T. gondii, respectively
(Vikoren et al., 2004; Gaffuri et al., 2006; Gamarra et al.,
2008). Between 8% and 38% of the wild boars tested in Eur-
ope were seropositive for T. gondii (Lutz, 1997; Gauss et
al., 2005; Bartova et al., 2006; Antolova et al., 2007). It
was recently shown that 22% of Western Grey Kangaroos
are T. gondii seropositive (Parameswaran, N., 2007. Devel-
opment of an ELISA for the detection of T. gondii antibod-
ies in macropod marsupials. In, WAAVP, Gent). Earlier
studies have indicated that kangaroos are highly suscepti-
ble to T. gondii infection and that the meat of infected ani-
mals contains viable tissue cysts (Canfield et al., 1990).
Kangaroo meat was implicated as the cause of an outbreak
of toxoplasmosis in Australia in 1994 (Robson et al., 1995).
4. Mixed meat products
Meat products are often made by mixing the meat and
organs of many animals. Many by-products such as sau-
sages or ham include pork meat. It has been estimated that
one pig may be consumed by 200–400 individuals
(Fehlhaber et al., 2002). Older animals (sows) are mainly
used for cured meat products and it is known that T. gondii
infection increases with age (Tenter et al., 2000). In the
event of including a few infected animals, a whole batch
may become infected (Aspinall et al., 2002). The actual
presence of viable T. gondii organisms in commercially
made fresh pork sausages was recently shown in a study
from Brazil (Dias et al., 2005). Viable parasites were also
detected in one out of 67 cured meat samples investigated
in the United Kingdom (Warnekulasuriya et al., 1998).
Mixture of beef with other types of meat is general practice
in minced meat preparations and could be an explanation
for the observed epidemiological findings that link con-
sumption of raw beef to T. gondii infection in humans.
5. Toxoplasma gondii ‘‘on farm risk’’ management
5.1. Sources of infection
To prevent T. gondii infection on the farm it is necessary
to identify the sources of infection in relation to the behav-
iour of various farm animals. Herbivorous animals most
likely contract T. gondii infection via pasture, hay, forage,
feed or surface water infected with oocysts shed by infected
cats (Skjerve et al., 1998; Tenter et al., 2000). Control of the
presence of cats on the farm or the shedding of oocysts
from these cats may play an important role in T. gondii pre-
vention of herbivorous farm animals (Frenkel et al., 1991;
Mateus-Pinilla et al., 2002) (Fig. 1). As yet little data is
available concerning the analysis of risk factors for herbiv-
orous animals to become infected with T. gondii.
Omnivorous animals can become infected with T. gondii
similar to herbivorous animals, but also via uptake of
rodents carrying T. gondii cysts (Fig. 1). In particular, this
may be a risk factor for poultry or pigs (Weigel et al., 1995;
Kijlstra et al., 2008). Feeding of non-pasteurized goat whey
has been identified as a risk factor for T. gondii infection in
pigs (Meerburg et al., 2006), and T. gondii DNA has
recently been shown to be present in sheep milk (Fusco
et al., 2007). Feeding of raw milk products to other animals
(and humans) might also play a role in the horizontal
transmission of T. gondii infection (Fusco et al., 2007).
The relative role of oocysts versus tissue cysts in T. gondii
infection of omnivorous farm animals such as pigs has
not yet been elucidated.
5.2. Cat control
Cats with outdoor access may become infected via
oocysts in the environment or by predating wildlife
(Hejlicek and Literak, 1998; Woods et al., 2003; Afonso
et al., 2006). Primary infection in cats protected these ani-
mals from oocyst reshedding up to 6 years post primary
infection, under experimental conditions (Davis and
Dubey, 1995; Dubey, 1995). Reshedding of oocysts has
been observed after superinfection with other coccidia or
after immunosuppression under experimental conditions
(Dubey, 1995; Tenter et al., 2000). No evidence is available
whether naturally infected cats will reshed T. gondii oocysts
later in life.
An oral vaccine composed of live bradyzoites from an
oocyst-negative mutant strain (T-263) has been effective
in preventing oocyst shedding by cats (Frenkel et al.,
1991). The duration of the protective immunity has not
been established and as yet this vaccine is not commercially
available. A study investigating the effect of T-263 vaccina-
tion of cats on a farm, showed a moderate decrease in sero-
prevalence of farm rodents and pigs (Mateus-Pinilla et al.,
 A. Kijlstra, E. Jongert / International Journal for Parasitology 38 (2008) 1359–1370 1363

Fig. 1. Pre- and post-harvest risk management for the control of infectious sources of animal and human toxoplasmosis. (1) Freezing and/or heating. (2)
Heating of milk products. (3) Animal friendly production system. (4) Indoor production system. (5) Cat control. (6) Rodent control. (7) Animal feed &
bedding decontamination by heating. (8) Vaccination against tissue cyst formation (a) or oocyst shedding (b). (9) Serological monitoring of people at risk.
(10) Consumer education on food hygiene and decontamination procedures. Percentages indicate highest observed seroprevalence per species. Thickness
of arrows represents the likeliness of transmission without risk management procedure. Percentages indicate the highest observed seroprevalence per
animal species or the percentage of cats actively shedding oocysts.

1999). In view of the long-term oocyst survival and vertical
transfer of infection in intermediate hosts (Marshall et al.,
2004), the effect of vaccination of cats on T. gondii seropre-
valence in pigs may take several years (Mateus-Pinilla et
al., 2002). Recently, it was shown that intranasal vaccina-
tion of cats with crude rhoptry proteins in immune stimu-
lating complexes (ISCOMs) prevented oocyst shedding in
two out of three challenged cats (Garcia et al., 2007).
Although a farmer can vaccinate his own cats against
T. gondii oocyst shedding, this will not prevent contamina-
tion of the farm area by non-vaccinated stray or neighbour
cats. It remains questionable whether cat owners can be
persuaded to participate in general T. gondii vaccination
programs that have a long-term food safety objective, but
do not contribute towards the health status of individual
cats. A successful approach in T. gondii vaccination of cats
may therefore depend on government regulations. In order
to reduce the presence of T. gondii oocysts on a farm, the
presence of cats on the farm should be avoided.
5.3. Farm animal vaccination
Toxoplasma gondii vaccination of farm animals is tra-
ditionally used to prevent abortions in sheep and goats.
A commercially available tachyzoite vaccine (OvilisÒ
Toxovax, Intervet) consisting of a live mutant strain
(S48) that does not persist in sheep tissues is used in
countries where parasite associated abortions are fre-
quently encountered (Innes and Vermeulen, 2006). How-
ever, vaccination with Toxovax does not eliminate
vertical transmission of the parasite when infection
occurs during pregnancy. Vaccination studies in sheep
with a live non-persistent T. gondii strain (RH) encapsu-
lated in ISCOMs or poly-lactide-co-glycolide (PLG)
microspheres could induce increased immune responses
after challenge, but did not result in significant protec-
tion (Buxton et al., 1989). Vaccination of pigs with the
live RH strain could reduce parasite load in the tissues
of these animals (Kotula et al., 1991), resulting in T.
gondii-free meat in 52% of vaccinated pigs after oral
challenge with T. gondii oocysts (Kringel et al., 2004).
Challenge of pigs vaccinated with a T. gondii rhoptry
extract failed to protect animals against experimental
acute toxoplasmosis, however 37% of surviving animals
were free of tissue cysts (Garcia et al., 2007). Recently,
a cocktail DNA vaccine has been shown to prime the
immune system of pigs against toxoplasmosis; increased
immune responses were observed after experimental chal-
lenge, resulting in Toxoplasma-free meat in two of three
vaccinated animals (Jongert et al., 2008).
1364 A. Kijlstra, E. Jongert / International Journal for Parasitology 38 (2008) 1359–1370
From a food safety perspective, vaccination could be
employed to reduce or prevent the formation of tissue cysts
in meat (Fig. 1).
5.4. Farm management
In the case of environmental contamination with
oocysts the most extreme management measure would
be to keep farm animals intended for meat production
inside, heat all feed administered to a temperature of at
least 70 °C and provide animals with clean drinking
water. To prevent oocyst contamination of food or stables
it is important to keep cats out of these areas. For those
food animals that can become infected via uptake of tis-
sue cysts it is important to prevent access of rodents
and birds into the stables, bedding material and forage
(Kijlstra et al., 2004b). In the case of free range farming
it is necessary to take adequate measures to control
rodents in the vicinity of the stables (Meerburg et al.,
2004), and feeding of farm animals with raw milk or milk
products (whey) from goats or sheep should be discour-
aged (Meerburg et al., 2006). Recently, rodent control
campaigns were shown to have an important impact on
the production of Toxoplasma-free pigs on organic farms
(Kijlstra et al., 2008).
6. Post-harvest prevention
6.1. Monitoring
Monitoring animals at slaughter would enable the iden-
tification of farms at risk and could be used to implement
changes in farm management to improve the food safety
of the meat produced. On the other hand it could be used
to detect T. gondii-infected meat and destroy the tissue
cysts in the meat by specific procedures (Fig. 1). Despite
the fact that the disease burden of toxoplasmosis is proba-
bly as high as salmonellosis and campylobacteriosis
(Kemmeren et al., 2006; Havelaar et al., 2007), T. gondii
in food animals is not monitored at slaughter whereas Sal-
monella and Campylobacter monitoring programs have
started in many countries. One of the problems with T.
gondii monitoring in food animals is the fact that there is
no general agreement among the tests to be used; no stan-
dardized reference sera or other reference materials are
available and there is no laboratory certification program.
A separate problem is the importation of fresh meat,
whereby the T. gondii status of the original animals is not
known.
6.2. Meat Handling, consumption and consumer attitude
Knowledge of consumers about toxoplasmosis is limited
and often restricted to women who have been informed
about the problem during pregnancy. Prevention pro-
grammes have been shown to be very efficient in reducing
the incidence of CT. A survey of 22 years of prevention
showed that seroconversion rates during pregnancy could
be reduced from 1.43% to 0.09% (Breugelmans et al.,
2004). In view of the fact that recent studies have shown
that the disease burden of acquired toxoplasmosis is also
high, it should become evident that public health informa-
tion concerning prevention of toxoplasmosis should be
directed at the public at large.
Nibbling of minced meat during food preparation has
been shown to be a risk factor for contracting toxoplasmo-
sis (Kapperud et al., 1996). The same study also showed
that women who washed kitchen knifes infrequently after
cutting meat also had an increased risk of infection, sug-
gesting cross-contamination as a mechanism of transmis-
sion. An outbreak of toxoplasmosis in Northern
Canada’s Inuit population was ascribed to the skinning
of fur and consumption of raw caribou meat from hunted
game, which shows that handling of infected animals may
be a risk for infection (McDonald et al., 1990).
As there is a large variation in the types and amounts of
meat eaten, and the way meats and meat products are pre-
pared throughout the world, each country should make
their own risk assessment and based on the findings imple-
ment management options for efficient prevention of T.
gondii infection via meat.
7. Killing the parasite
Studies have indicated that T. gondii tissue cysts in meat
are susceptible to various physical procedures such as heat
treatment, freezing, irradiation, high-pressure, acidity and
enhancing solutions. It should be noted that most studies
were performed using experimentally infected pigs or with
tissue cysts isolated from the brains of infected mice. Few
studies have evaluated effects on meat from naturally
infected animals. Heat treatment is the most secure method
to inactivate the parasite, whereby the responsibility of
food safety is laid in the hands of the consumer. Post-har-
vest but pre-kitchen procedures involving a combination of
treatments may be implemented to assure non-infectivity of
meat. In this respect a freezing procedure would be the
most practical risk management option.
7.1. Early studies
The hypothesis of horizontal transmission of T. gondii
by ingestion of undercooked meat was first proposed in
the late 1950’s (Weinman and Chandler, 1956), and tech-
niques to destroy the cyst form of the parasite were inves-
tigated (Table 2). The first experiments describing the
inactivation of tissue cysts of T. gondii examined the effects
of storage conditions on parasite survival and showed that
parasite tissue cysts could be lysed in distilled water (Jacobs
et al., 1960), but survived for several weeks in the presence
of physiological saline (0.85%) and storage at 4 °C (Jacobs
et al., 1960) (Dubey et al., 1990). Raising the salt concen-
tration or the temperature led to inactivation of the para-
site (Table 2).
 A. Kijlstra, E. Jongert / International Journal for Parasitology 38 (2008) 1359–1370 1365

Table 2
Effect of salting, freezing and heating on viability of Toxoplasma gondii tissue cysts
Sample Salt Temperature (°C) Time Efficacya Reference
Tissue cysts from infected rat brain 0.85% 4 0–24 days Jacobs et al. (1960)
0.1% 18–20 12 h +
0% 18–20 30 min +
0.1–2.1% 18–20 24 h +
0.85% 50 1h +
0.85% 56 10 min +
0.85% 50 15 min +
Tissue cysts from infected mouse brain 6% Various + Dubey (1997)
0.85% 56 days
2% 49 days
3.3% 21 days
2% 4 7 days + Hill et al. (2004)
1% 4 45 days
20 3 days + Djurkovic-Djakovic and
Milenkovic (2000)
Pork meat spiked with Toxoplasma cysts 2% 4 7 days + Hill et al. (2004)
1% 4 45 days
67 immediate + Dubey et al. (1990)
20 2 days + Sommer et al. (1965)
15% NaNO3/NaCl 5 4–21 days +c Scupin (1968)
12 + Kotula et al. (1991)
Various enhancing solutions 4 8h + Hill et al. (2006)
Meat from experimentally infected pigs 25 6–35 days +b Grossklaus and Baumgarten (1968)
Various enhancing solutions 4 8h + Hill et al. (2006)
7 to 12 Kuticic and Wikerhauser (1996)
Meat from infected sheep Salt & sugar 4 64 h + Lunden and Uggla (1992)
Smoking <50 24–48 h +
a
Efficacy was scored as or +; indicates that the procedure did not kill all parasites and + indicates a 100% killing effect of the procedure on the
parasite, unless stated otherwise. Parasite viability was evaluated by mice or cat bioassay.
b
One of 54 deep frozen samples was still positive in a mouse bioassay after storage at 25 °C for 35 days.
c
Toxoplasma gondii cysts could be isolated up to 13 days after preparation of smoked hams.

7.2. Freezing and heating
The effect of freezing on T. gondii cyst viability was first
described in 1965 (Sommer et al., 1965). It was observed
that freezing for 2 days at 20 °C was sufficient to inacti-
vate the parasite. Experiments with meat from pigs that
were fed with T. gondii-infected mice, showed that all meat
samples were rendered non-infectious by freezing 6–35
days at 25 °C (Grossklaus and Baumgarten, 1968).
Experiments using different freezing temperatures showed
that an internal temperature of 12 °C was sufficient to
render the parasite non-viable (Kotula et al., 1991), and
that parasites in meat from experimentally infected pigs
did not survive when frozen for 4 days at 7 °C to
12 °C (Kuticic and Wikerhauser, 1996). Other studies
showed that at least 3 days at 20 °C were required to
inactivate isolated tissue cysts (Djurkovic-Djakovic and
Milenkovic, 2000). Toxoplasma gondii tissue cysts
remained viable up to 22 days at 1 and 3.9 °C and 11
days at 6.7 °C (Kotula et al., 1991). This latter study thus
indicates that importation of marginally frozen meats does
not guarantee destruction of the parasite. In general, freez-
ing can inactivate the T. gondii tissue cysts, but proper tim-
ing and temperature are necessary for a 100% parasite
killing efficiency. Freezing of meat by consumers is widely
applied in westernized countries. The loss of sensory qual-
ity may be an important factor in consumers’ attitudes
towards freezing of meat.
Jacobs et al. (1960) were the first to show that heating
could inactivate tissue cysts. At 50 °C it takes 1 h to inacti-
vate tissue cysts (Table 2) whereas immediate destruction
takes place when the internal temperature of meat reaches
a temperature of 67 °C (Dubey et al., 1990). Cooking
infected meat in a microwave does not guarantee killing
of the parasite, most probably due to uneven heating
(Lunden and Uggla, 1992). The primary control factor
for prevention of T. gondii infection via meat consumption
is adequate cooking and prevention of cross-contamination
(McCurdy et al., 2006). Limited data are available concern-
ing consumer cooking habits and it is certainly possible
that parts of meat being grilled or barbecued do not reach
sufficiently high-temperatures to kill the parasite. Cooking
style has an influence on cooking temperatures and time,
and has been shown to have a considerable impact on
the reduction of Escherichia coli contamination in ground
beef (Berry and Bigner-George, 2001; Rhee et al., 2003).
1366 A. Kijlstra, E. Jongert / International Journal for Parasitology 38 (2008) 1359–1370
7.3. Pumping and curing meat
A number of studies have addressed the effect of curing
procedures such as salting, smoking or fermentation on tis-
sue cyst survival. Curing of meat has historically been
introduced to preserve meat. Curing of lamb meat with salt
and sugar for 64 h at 4 °C or smoking salt-injected meat at
temperatures not exceeding 50 °C for 24–28 h were effective
in killing T. gondii (Lunden and Uggla, 1992). Studies by
Dubey have shown that 6% NaCl can kill the isolated tissue
cyst, independent of the temperature used (Dubey, 1997).
Isolated tissue cysts can survive for 56 days in a solution
of 0.85% salt, 49 days at 2% and 21 days at 3.3% (Dubey,
1997). More recent data have shown that injection of >2%
NaCl and/or >1.4% lactate salt solutions into experimen-
tally infected pig meat could kill the parasite but that a
1% NaCl solution provided variable results, while the addi-
tion of tripolyphosphate salts had no effect on parasite via-
bility (Hill et al., 2004, 2006). It should be noted that the
above experiments were performed with the VEG strain
and that the effect of strain differences with regard to via-
bility following treatment with enhancing solutions has
not been investigated yet.
Curing of meat products often involves the mixing of
meat from various animals from different farms and some-
times from different farming systems (organic and regular),
and thus a few infected animals may lead to T. gondii con-
tamination of a whole batch of cured meat products. PCR
studies have demonstrated the presence of T. gondii DNA
in commercially available swine sausages but bio-assays
could not detect the presence of viable tissue cysts in these
samples (de Oliveira Mendonca et al., 2004). However, via-
ble T. gondii cysts were detected in freshly prepared swine
sausages in other experiments (Dias et al., 2005). The differ-
ence between these two studies may be ascribed to the time
interval between sausage preparation and bioassay in mice.
Other studies indicated that depending upon the time of
incubation, salt used for preparing sausages may inactivate
T. gondii (Jamra et al., 1991). The high prevalence of toxo-
plasmosis in certain areas of the world has been associated
with the consumption of raw sausages (Buffolano et al.,
1996; Dias et al., 2005).
7.4. Irradiation and high-pressure
Gamma irradiation of tissue cysts can render the para-
site inactive at doses between 0.4 and 0.7 kGy (Dubey et
al., 1986; Dubey and Thayer, 1994; Kuticic and
Wikerhauser, 1996). However, the adverse effects of irradi-
ation on colour have a major impact on the use of this tech-
nology and in certain countries large-scale implementation
is restricted due to poor consumer acceptance (Brewer,
2004; Aymerich et al., 2008).
High-pressure treatment using 300 MPa can inactivate
T. gondii tissue cysts (Lindsay et al., 2006) but negative
effects on meat colour and texture have to be addressed
before this method can be developed for routine decontam-
ination (Cheftel and Culioli, 1997).
8. Conclusion
Toxoplasma gondii is a ubiquitous parasite that can
infect almost any warm-blooded animal. The parasite has
a complex life cycle and multiple routes of infection are
possible. New developments in the knowledge of T. gondii
infection, the associated disease burden and the current
inability to pharmacologically kill the tissue cyst stage of
the parasite demand stricter monitoring and surveillance
of the infection in both humans and food animals.
In this report, we review the data implicating meat as an
important source of infection and describe strategies to
reduce the risk of human T. gondii infection via this source
(Fig. 1). Infection in humans usually occurs via the con-
sumption of cysts in undercooked meat (products) or by
the uptake of soil or water contaminated by oocysts from
feline faeces. The relative role of these routes of transmis-
sion are not exactly known since disease manifestations
cannot be distinguished according to source of infection,
and no laboratory tests are available to distinguish an
infection by oocysts from an infection by tissue cysts.
The relative role of infection sources therefore comes from
epidemiological studies. Epidemiological case-control stud-
ies have attributed the main source of infection in Europe
to undercooked meat. Meat from grazing animals such as
sheep and goats can be considered as important potential
sources of T. gondii infection for humans. Consumption
of horse, game and chicken can also be considered to play
a role in horizontal transmission of T. gondii to humans.
Despite epidemiological studies pointing to beef as a source
of T. gondii infection in humans, viable parasites have not
been isolated from commercial cuts of beef meat. This may
be due to confounding factors in epidemiological studies
such as general meat knowledge of the respondents, in par-
ticular on the presence of other meat sources in products
sold as beef (adulteration) and meat mixtures where beef
is a component amongst others.
Depending on culture, religion, and food preparation
methods, different sources of meat may act as most impor-
tant infection source for a certain population, indicating
that prevention needs to be tailored according to social
habits per country or region of the world. Although the rel-
ative role of tissue cysts versus oocysts in human toxoplas-
mosis is not exactly clear and may differ between countries,
the former can be localized and inactivated whereas pre-
vention of the latter source is more complex (Dubey,
2004; Sroka et al., 2006). Toxoplasmosis in humans, caused
by the consumption of infected meat, is a preventable dis-
ease entity. Preventing infection of food animals can be
achieved by giving animals sterilized feed and water, keep-
ing animals indoors and preventing access of other ani-
mals. Modern production technologies have shown that
this is feasible and have led to a marked decrease of T. gon-
dii infections in meat producing animals such as pigs. Pub-
 A. Kijlstra, E. Jongert / International Journal for Parasitology 38 (2008) 1359–1370
lic demand for animal friendly production systems may
however lead to a re-emergence of T. gondii in pork and
poultry. Other potential sources of infection such as goat,
sheep, horses and game have remained relatively
unchanged and their role in human infection depends on
the degree of exposure, post-harvest treatment (freezing)
and consumer kitchen and eating habits.
Reduction of the environmental oocyst load is probably
the most important management practice to prevent trans-
fer to farm animals. As long as no commercial vaccines are
available to prevent oocyst shedding by cats or prevent tis-
sue cysts in meat animals, other preventive measures can be
taken. As an extreme, farms should not be allowed to have
cats, or at least the number of cats on the farm should be
limited, and should have no access to stables and feed sup-
ply. Control of farm cats should be included in Quality
Assurance programs. A monitoring and surveillance sys-
tem should be set up for animal friendly production sys-
tems. Monitoring at slaughter is currently hampered by
the absence of standardized serological methods. Follow-
ing implementation of standardized T. gondii monitoring
it is recommended that all animals with a seropositive sta-
tus should be decontaminated after slaughter. The produc-
tion of T. gondii-safe meat is technically feasible and can be
achieved by post-harvest decontamination procedures.
Evaluation of decontamination procedures indicates that
freezing is the best option to produce T. gondii-safe meat,
although details of the procedure need to be validated.
Other methods such as irradiation do not share consumer
approval and high-pressure disinfection methods need to
be developed further for large-scale use. Pre-harvest risk
management of game in relation to T. gondii is inherently
impossible.
There is a necessity to develop novel risk management
options to prevent transmission from meat to humans,
including better consumer education, slaughterhouse mon-
itoring, improved farm management, post-harvest decon-
tamination or development of a vaccine against human
toxoplasmosis. Consumer education has traditionally been
directed to pregnant women and shown to be successful
(Breugelmans et al., 2004). In view of recent developments
in the epidemiology of the disease and calculations of dis-
ease burden it is clear that not only pregnant women
should be addressed but that the whole population is at
risk of adverse effects of the disease and should be informed
how to prevent infection. Ultimately, a vaccine against
toxoplasmosis may prevent human infection and/or
pathology, irrespective of the source of contamination.
Until then, consumers should be informed about the risks
of T. gondii infections associated with meat and meat prod-
ucts and could be advised to freeze meat a few days before
cooking to ensure higher inactivation efficiency.
Acknowledgements
This study was supported by the Dutch Ministry of
Agriculture, Nature and Food Quality and the Belgian
1367
Federal Public Service for Health, Food Chain Safety
and Environment. We acknowledge the a.s.b.l. ‘‘Les amis
de l’Institut Pasteur de Bruxelles” for their support.
References
Afonso, E., Thulliez, P., Gilot-Fromont, E., 2006. Transmission of
Toxoplasma gondii in an urban population of domestic cats (Felis
catus). Int. J. Parasitol. 36, 1373–1382.
Afonso, E., Lemoine, M., Poulle, M.L., Ravat, M.C., Romand, S.,
Thulliez, P., Villena, I., Aubert, D., Rabilloud, M., Riche, B., Gilot-
Fromont, E., 2008. Spatial distribution of soil contamination by
Toxoplasma gondii in relation to cat defecation behaviour in an urban
area. Int. J. Parasitol. 38, 1017–1023.
AFSSA, 2005. Toxoplasmose:état des connaisances et évaluation du risque
lié à l’alimentation. In: Report of the Afssa working group ‘‘Toxoplasma
gondii”. http://www.afssa.fr/Documents/MIC-Ra-Toxoplasmose.pdf.
Alkhalidi, N.W., Dubey, J.P., 1979. Prevalence of Toxoplasma gondii
infection in horses. J. Parasitol. 65, 331–334.
Antolova, D., Reiterova, K., Dubinsky, P., 2007. Seroprevalence of
Toxoplasma gondii in wild boars (Sus scrofa) in the Slovak Republic.
Ann. Agric. Environ. Med. 14, 71–73.
Aspinall, T.V., Marlee, D., Hyde, J.E., Sims, P.F.G., 2002. Prevalence of
Toxoplasma gondii in commercial meat products as monitored by
polymerase chain reaction – food for thought? Int. J. Parasitol. 32,
1193–1199.
Aymerich, T., Picouet, P.A., Monfort, J.M., 2008. Decontamination
technologies for meat products. Meat Sci. 78, 114–129.
Bahia-Oliveira, L.M.G., Jones, J.L., Azevedo-Silva, J., Alves, C.C.F.,
Orefice, F., Addiss, D.G., 2003. Highly endemic, waterborne
toxoplasmosis in north Rio de Janeiro state. Brazil Emerg. Infect.
Dis. 9, 55–62.
Baril, L., Ancelle, T., Goulet, V., Thulliez, P., Tirard-Fleury, V., Carme,
B., 1999. Risk factors for Toxoplasma infection in pregnancy: a case-
control study in France. Scand. J. Infect. Dis. 31, 305–309.
Bartova, E., Sedlak, K., Literak, I., 2006. Prevalence of Toxoplasma gondii
and Neospora caninum antibodies in wild boars in the Czech Republic.
Vet. Parasitol. 142, 150–153.
Berry, B.W., Bigner-George, M.E., 2001. Postcooking temperature
changes in beef patties. J. Food Prot. 64, 1405–1411.
Blewett, D.A., Miller, J.K., Buxton, D., 1982. Response of immune and
susceptible ewes to infection with Toxoplasma gondii. Vet. Rec. 111, 175–
177.
Bottieau, E., Clerinx, J., Van den Enden, E., Van Esbroeck, M.,
Colebunders, R., Van Gompel, A., Van den Ende, J., 2006. Infectious
mononucleosis-like syndromes in febrile travelers returning from the
tropics. J. Travel Med. 13, 191–197.
Bowie, W.R., King, A.S., Werker, D.H., IsaacRenton, J.L., Bell, A., Eng,
S.B., Marion, S.A., 1997. Outbreak of toxoplasmosis associated with
municipal drinking water. Lancet 350, 173–177.
Breugelmans, M., Naessens, A., Foulon, W., 2004. Prevention of
toxoplasmosis during pregnancy – an epidemiologic survey over 22
consecutive years. J. Perinat. Med. 32, 211–214.
Brewer, S., 2004. Irradiation effects on meat color – a review. Meat Sci. 68,
1–17.
Buffolano, W., Gilbert, R.E., Holland, F.J., Fratta, D., Palumbo, F.,
Ades, A.E., 1996. Risk factors for recent Toxoplasma infection in
pregnant women in Naples. Epidemiol. Infect. 116, 347–351.
Buxton, D., Uggla, A., Lovgren, K., Thomson, K., Lunden, A., Morein,
B., Blewett, D.A., 1989. Trial of a novel experimental Toxoplasma
iscom vaccine in pregnant sheep. Br. Vet. J. 145, 451–457.
Buzby, J.C., Roberts, T., 1996. ERS updates US foodborne diseases costs
for seven pathogens. Food Rev. 19, 20–25.
Canfield, P.J., Hartley, W.J., Dubey, J.P., 1990. Lesions of toxoplasmosis
in Australian marsupials. J. Comp. Pathol. 103, 159–167.
Carme, B., Bissuel, F., Ajzenberg, D., Bouyne, R., Aznar, C., Demar, M.,
Bichat, S., Louvel, D., Bourbigot, A.M., Peneau, C., Neron, P., Darde,
1368 A. Kijlstra, E. Jongert / International Journal for Parasitology 38 (2008) 1359–1370
M.L., 2002. Severe acquired toxoplasmosis in immunocompetent adult
patients in French Guiana. J. Clin. Microbiol. 40, 4037–4044.
Cheftel, J.C., Culioli, J., 1997. Effects of high pressure on meat: a review.
Meat Sci. 46, 211–236.
Childs, J.E., Seegar, W.S., 1986. Epidemiologic observations on infection
with Toxoplasma gondii in 3 species of urban mammals from
Baltimore, Maryland, USA. Int. J. Zoonoses 13, 249–261.
Choi, W.Y., Nam, H.W., Kwak, N.H., Huh, W., Kim, Y.R., Kang, M.W.,
Cho, S.Y., Dubey, J.P., 1997. Foodborne outbreaks of human
toxoplasmosis. J. Infect. Dis. 175, 1280–1282.
Cook, A.J.C., Gilbert, R.E., Buffolano, W., Zufferey, J., Petersen, E.,
Jenum, P.A., Foulon, W., Semprini, A.E., Dunn, D.T., 2000. Sources
of Toxoplasma infection in pregnant women: european multicentre
case-control study. Br. Med. J. 321, 142–147.
da Silva, D.S., Bahia-Oliveira, L.M.G., Shen, S.K., Kwok, O.C.H.,
Lehman, T., Dubey, J.P., 2003. Prevalence of Toxoplasma gondii in
chickens from an area in southern Brazil highly endemic to humans. J.
Parasitol. 89, 394–396.
Dabritz, H.A., Miller, M.A., Atwill, E.R., Gardner, I.A., Leutenegger,
C.M., Melli, A.C., Conrad, P.A., 2007. Detection of Toxoplasma
gondii-like oocysts in cat feces and estimates of the environmental
oocyst burden. J. Am. Vet. Med. Assoc. 231, 1676–1684.
Davies, P.R., Morrow, W.E.M., Deen, J., Gamble, H.R., Patton, S., 1998.
Seroprevalence of Toxoplasma gondii and Trichinella spiralis in
finishing swine raised in different production systems in North
Carolina, USA. Prev. Vet. Med. 36, 67–76.
Davis, S.W., Dubey, J.P., 1995. Mediation of immunity to Toxoplasma
gondii oocyst shedding in cats. J. Parasitol. 81, 882–886.
de Oliveira Mendonca, A., Domingues, P.F., da Silva, A.V., Pezerico,
S.B., Langoni, H., 2004. Detection of Toxoplasma gondii in swine
sausages. Parasitol. Latinoam. 59, 42–45.
Dias, R.A.F., Navarro, I.T., Ruffolo, B.B., Bugni, F.M., de Castro, M.V.,
Freire, R.L., 2005. Toxoplasma gondii in fresh pork sausage and
seroprevalence in butchers from factories in Londrina, Parana State,
Brazil(1). Rev. Inst. Med. trop. S. Paulo 47, 185–189.
Diza, E., Frantzidou, F., Souliou, E., Arvanitidou, M., Gioula, G.,
Antoniadis, A., 2005. Seroprevalence of Toxoplasma gondii in northern
Greece during the last 20 years. Clin. Microbiol. Infec. 11, 719–723.
Djurkovic-Djakovic, O., Milenkovic, V., 2000. Effect of refrigeration and
freezing on survival of Toxoplasma gondii tissue cysts. Acta Vet.
Beograd 50, 375–380.
Dubey, J.P., 1980. Mouse pathogenicity of Toxoplasma gondii isolated
from a goat. Am. J. Vet. Res. 41, 427–429.
Dubey, J.P., Sharma, S.P., Lopes, C.W.G., Williams, J.F., Williams,
C.S.F., Weisbrode, S.E., 1980. Caprine toxoplasmosis – abortion,
clinical signs, and distribution of Toxoplasma in tissues of goats fed
Toxoplasma gondii oocysts. Am. J. Vet. Res. 41, 1072–1076.
Dubey, J.P., 1981. Epizootic toxoplasmosis associated with abortion in
dairy goats in montana. J. Am. Vet. Med. Assoc. 178, 661–670.
Dubey, J.P., 1983. Distribution of cysts and tachyzoites in calves and
pregnant cows inoculated with Toxoplasma gondii oocysts. Vet.
Parasitol. 13, 199–211.
Dubey, J.P., Murrell, K.D., Fayer, R., 1984. Persistence of encysted
Toxoplasma gondii in tissues of pigs fed oocysts. Am. J. Vet. Res. 45,
1941–1943.
Dubey, J.P., 1986a. A review of toxoplasmosis in pigs. Vet. Parasitol. 19,
181–223.
Dubey, J.P., 1986b. A review of toxoplasmosis in cattle. Vet. Parasitol. 22,
177–202.
Dubey, J.P., Brake, R.J., Murrell, K.D., Fayer, R., 1986. Effect of
irradiation on the viability of Toxoplasma gondii cysts in tissues of mice
and pigs. Am. J. Vet. Res. 47, 518–522.
Dubey, J.P., 1988. Long-term persistence of Toxoplasma gondii in tissues
of pigs inoculated with T. gondii oocysts and effect of freezing on
viability of tissue cysts in pork. Am. J. Vet. Res. 49, 910–913.
Dubey, J.P., Kotula, A.W., Sharar, A., Andrews, C.D., Lindsay, D.S.,
1990. Effect of high-temperature on infectivity of Toxoplasma gondii
tissue cysts in pork. J. Parasitol. 76, 201–204.
Dubey, J.P., Thulliez, P., 1993. Persistence of tissue cysts in edible
tissues of cattle fed Toxoplasma gondii oocysts. Am. J. Vet. Res. 54,
270–273.
Dubey, J.P., Thayer, D.W., 1994. Killing of different strains of Toxo-
plasma gondii tissue cysts by irradiation under defined conditions. J.
Parasitol. 80, 764–767.
Dubey, J.P., 1995. Duration of immunity to shedding of Toxoplasma
gondii oocysts by cats. J. Parasitol. 81, 410–415.
Dubey, J.P., 1996. Strategies to reduce transmission of Toxoplasma gondii
to animals and humans. Vet. Parasitol. 64, 65–70.
Dubey, J.P., 1997. Survival of Toxoplasma gondii tissue cysts in 0.85–6%
NaCl solutions at 4–20 °C. J. Parasitol. 83, 946–949.
Dubey, J.P., 2004. Toxoplasmosis – a waterborne zoonosis. Vet. Parasitol.
126, 57–72.
Dubey, J.P., Salant, H., Sreekumar, C., Dahl, E., Vianna, M.C.B., Shen,
S.K., Kwok, O.C.H., Spira, D., Hamburger, J., Lehmann, T.V., 2004.
High prevalence of Toxoplasma gondii in a commercial flock of
chickens in Israel, and public health implications of free-range
farming. Vet. Parasitol. 121, 317–322.
Dubey, J.P., Jones, J.L., 2008. Toxoplasma gondii infection in humans and
animals in the United States. Int. J. Parasitol. 38, 1257–1278.
EFSA, 2007. Monitoring of Toxoplasma in humans, food and animals.
EFSA Journal 583, 1–64.
Esteban-Redondo, I., Maley, S.W., Thomson, K., Nicoll, S., Wright, S.,
Buxton, D., Innes, E.A., 1999. Detection of T. Gondii in tissues of
sheep and cattle following oral infection. Vet. Parasitol. 86, 155–171.
Fehlhaber, K., Hintersdorf, P., Kruger, G., 2002. Study on the prevalence
of Toxoplasma gondii in pigs of different management systems and in
minced meat. Fleischwirtschaft 83, 97–99.
Frenkel, J.K., Pfefferkorn, E.R., Smith, D.D., Fishback, J.L., 1991.
Prospective vaccine prepared from a new mutant of Toxoplasma gondii
for use in cats. Am. J. Vet. Res. 52, 759–763.
Fusco, G., Rinaldi, L., Guarino, A., Proroga, Y., Pesce, A., Giuseppina
de, M., Cringoli, G., 2007. Toxoplasma gondii in sheep from the
Campania region (Italy). Vet. Parasitol. 149, 271–274.
Gaffuri, A., Giacometti, M., Tranquillo, V.M., Magnino, S., Cordioli, P.,
Lanfranchi, P., 2006. Serosurvey of roe deer, chamois and domestic
sheep in the central Italian Alps. J. Wildl. Dis. 42, 685–690.
Gamarra, J.A., Cabezon, O., Pabon, M., Arnal, M.C., Luco, D.F.,
Dubey, J.P., Gortazar, C., Almeria, S., 2008. Prevalence of antibodies
against Toxoplasma gondii in roe deer from Spain. Vet. Parasitol. 153,
152–156.
Garcia, J.L., Navarro, I.T., Biazzono, L., Freire, R.L., Guimaraes, J.D.,
Cryssafidis, A.L., Bugni, F.M., da Cunha, I.A.L., Hamada, F.N.,
Dias, R.C.F., 2007. Protective activity against oocyst shedding in cats
vaccinated with crude rhoptry proteins of the Toxoplasma gondii by
the intranasal route. Vet. Parasitol. 145, 197–206.
Gauss, C.B.L., Dubey, J.P., Vidal, D., Ruiz, F., Vicente, J., Marco, I.,
Lavin, S., Gortazar, C., Almeria, S., 2005. Seroprevalence of Toxo-
plasma gondii in wild pigs (Sus scrofa) from Spain. Vet. Parasitol. 131,
151–156.
Gilbert, R.E., Stanford, M.R., 2000. Is ocular toxoplasmosis caused by
prenatal or postnatal infection? Br. J. Ophthalmol. 84, 224–226.
Gill, C.O., 2005. Safety and storage stability of horse meat for human
consumption. Meat Sci. 71, 506–513.
Glasner, P.D., Silveira, C., Kruszonmoran, D., Martins, M.C., Burnier,
M., Silveira, S., Camargo, M.E., Nussenblatt, R.B., Kaslow, R.A.,
Belfort, R., 1992. An unusually high prevalence of ocular toxoplas-
mosis in southern Brazil. Am. J. Ophthalmol. 114, 136–144.
Gormley, P.D., Pavesio, C.E., Minnasian, D., Lightman, S., 1998. Effects
of drug therapy on Toxoplasma cysts in an animal model of acute and
chronic disease. Invest. Ophthalmol. Vis. Sci. 39, 1171–1175.
Grossklaus, D., Baumgarten, H.-J., 1968. Die überlebensdauer von
Toxoplasma-cysten in schweinefleisch I. Mitteilung: ergebnisse von
lagerungsversuchen bei verschiedenen temperaturen. Fleischwirtschaft
48, 930–932.
Hall, S.M., Pandit, A., Golwilkar, A., Williams, T.S., 1999. How do Jains
get toxoplasma infection? Lancet 354, 486–487.
 A. Kijlstra, E. Jongert / International Journal for Parasitology 38 (2008) 1359–1370
Havelaar, A.H., Kemmeren, J.M., Kortbeek, L.M., 2007. Disease burden
of congenital toxoplasmosis. Clin. Infect. Dis. 44, 1467–1474.
Hejlicek, K., Literak, I., 1998. Long-term study of Toxoplasma gondii
prevalence in small mammals (Insectivora and Rodentia). Folia Zool
47, 93–101.
Hellmann, E., Tauscher, L., 1967. Research into the occurrence of
Toxoplasma in fresh beef and pork. Berl. Munch. Tierarztl. Woc-
henschr. 80, 209–212.
Hill, A.E., Benedetto, S.M.C., Coss, C., McCrary, J.L., Fournet, V.M.,
Dubey, J.P., 2006. Effects of time and temperature on the viability of
Toxoplasma gondii tissue cysts in enhanced pork loin. J. Food Prot. 69,
1961–1965.
Hill, D.E., Sreekumar, C., Gamble, H.R., Dubey, J.P., 2004. Effect of
commonly used enhancement solutions on the viability of Toxoplasma
gondii tissue cysts in pork loin. J. Food Prot. 67, 2230–2233.
Holland, G.N., 2003. LX Edward Jackson memorial lecture – ocular
toxoplasmosis: a global reassessment. Part 1: epidemiology and course
of disease. Am. J. Ophthalmol. 136, 973–988.
Innes, E.A., Vermeulen, A.N., 2006. Vaccination as a control strategy
against the coccidial parasites Eimeria, Toxoplasma and Neospora.
Parasitology 133, S145–S168.
Jacobs, L., Remington, J.S., Melton, M.L., 1960. The resistance of the
encysted form of Toxoplasma gondii. J. Parasitol. 46, 11–21.
Jamra, L.M., Martins, M.C., Vieira, M.D.L., 1991. Action of table salt
on Toxoplasma gondii. Rev. Inst. Med. Trop. Sao Paulo 33, 359–
363.
Jones, J.L., Kruszon-Moran, D., Sanders-Lewis, K., Wilson, M., 2007.
Toxoplasma gondii infection in the United States, 1999–2004, decline
from the prior decade. Am. J. Trop. Med. Hyg. 77, 405–410.
Jongert, E., Melkebeek, V., De Craeye, S., Dewit, J., Verhelst, D., Cox, E.,
2008. An enhanced GRA1–GRA7 cocktail DNA vaccine primes anti-
Toxoplasma immune responses in pigs. Vaccine 26, 1025–1031.
Kapperud, G., Jenum, P.A., StrayPedersen, B., Melby, K.K., Eskild, A.,
Eng, J., 1996. Risk factors for Toxoplasma gondii infection in
pregnancy – results of a prospective case-control study in Norway.
Am. J. Epidemiol. 144, 405–412.
Kemmeren, M., Mangen, M.J.J., van Duynhoven, Y.T.H.P., Havelaar,
A.H. 2006. Priority setting of foodborne pathogens, disease burden
and costst of selected enteric pathogens. Bilthoven: National Institute
for Public Health and the Environment; Report nr. 330080001. http://
www.rivm.nl/bibliotheek/rapporten/330080001.pdf.
Kijlstra, A., Eissen, O.A., Cornelissen, J., Munniksma, K., Eijck, I.,
Kortbeek, T., 2004a. Toxoplasma gondii infection in animal-friendly
pig production systems. Invest. Ophthalmol. Vis. Sci. 45, 3165–3169.
Kijlstra, A., Meerburg, B.G., Mul, M.F., 2004b. Animal-friendly produc-
tion systems may cause re-emergence of Toxoplasma gondii. Njas-
Wageningen J. Life Sci. 52, 119–132.
Kijlstra, A., Meerburg, B., Cornelissen, J., De Craeye, S., Vereijken, P.,
Jongert, E., 2008. The role of rodents and shrews in the transmission of
Toxoplasma gondii to pigs. Vet. Parasitol., in press [Epub ahead of
print].
Kniel, K.E., Lindsay, D.S., Sumner, S.S., Hackney, C.R., Pierson, M.D.,
Dubey, J.P., 2002. Examination of attachment and survival of
Toxoplasma gondii oocysts on raspberries and blueberries. J. Parasitol.
88, 790–793.
Kortbeek, L.M., De Melker, H.E., Veldhuijzen, I.K., Conyn-Van
Spaendonck, M.A.E., 2004. Population-based toxoplasma seropre-
valence study in the Netherlands. Epidemiol. Infect. 132, 839–845.
Kotula, A.W., Dubey, J.P., Sharar, A.K., Andrews, C.D., Shen, S.K.,
Lindsay, D.S., 1991. Effect of freezing on infectivity of Toxoplasma
gondii tissue cysts in pork. J. Food Prot. 54, 687–690.
Kringel, H., Dubey, J.P., Beshah, E., Hecker, R., Urban, J.F., 2004. CpG-
oligodeoxynucleotides enhance porcine immunity to Toxoplasma
gondii. Vet. Parasitol. 123, 55–66.
Kuticic, V., Wikerhauser, T., 1996. Studies of the effect of various
treatments on the viability of Toxoplasma gondii tissue cysts and
oocysts, Toxoplasma gondii. Curr. Top. Microbiol. Immunol. 219,
261–265.
1369
Lake, R., Hudson, A., Cressey, P., 2002. Risk profile: Toxoplasma gondii
in red meat and meat products; http://www.nzfsa.govt.nz/science/risk-
profiles/Toxoplasmagondii-in-red-meat.pdf. In: New Zealand Food
Safety Authority, pp. 1–33.
Lehmann, T., Marcet, P.L., Graham, D.H., Dahl, E.R., Dubey, J.P., 2006.
Globalization and the population structure of Toxoplasma gondii.
Proc. Natl. Acad. Sci. USA 103, 11423–11428.
Lindsay, D.S., Collins, M.V., Holliman, D., Flick, G.J., Dubey, J.P., 2006.
Effects of high-pressure processing on Toxoplasma gondii tissue cysts in
ground pork. J. Parasitol. 92, 195–196.
Lunden, A., Uggla, A., 1992. Infectivity of Toxoplasma gondii in mutton
following curing, smoking, freezing or microwave cooking. Int. J.
Food Microbiol. 15, 357–363.
Lutz, W., 1997. Serological evidence of antibodies against Toxoplasma
and Leptospira in wild boar. Zeitschrift Fur Jagdwissenschaft 43, 283–
287.
Marshall, P.A., Hughes, J.M., Williams, R.H., Smith, J.E., Murphy, R.G.,
Hide, G., 2004. Detection of high levels of congenital transmission of
Toxoplasma gondii in natural urban populations of Mus domesticus.
Parasitology 128, 39–42.
Mateus-Pinilla, N.E., Dubey, J.P., Choromanski, L., Weigel, R.M., 1999.
A field trial of the effectiveness of a feline Toxoplasma gondii vaccine in
reducing T. Gondii exposure for swine. J. Parasitol. 85, 855–860.
Mateus-Pinilla, N.E., Hannon, B., Weigel, R.M., 2002. A computer
simulation of the prevention of the transmission of Toxoplasma gondii
on swine farms using a feline T-gondii vaccine. Prev. Vet. Med. 55, 17–
36.
McColgan, C., Buxton, D., Blewett, D.A., 1988. Titration of Toxoplasma
gondii oocysts in non-pregnant sheep and the effects of subsequent
challenge during pregnancy. Vet. Rec. 123, 467–470.
McCurdy, S.M., Takeuchi, M.T., Edwards, Z.M., Edlefsen, M., Kang,
D.H., Mayes, V.E., Hillers, V.N., 2006. Food safety education
initiative to increase consumer use of food thermometers in the United
States. Br. Food J. 108, 775–794.
McDonald, J.C., Gyorkos, T.W., Alberton, B., Maclean, J.D., Richer, G.,
Juranek, D., 1990. An outbreak of toxoplasmosis in pregnant-women
in Northern Quebec. J. Infect. Dis. 161, 769–774.
Mead, P.S., Slutsker, L., Dietz, V., McCaig, L.F., Bresee, J.S., Shapiro,
C., Griffin, P.M., Tauxe, R.V., 1999. Food-related illness and death in
the United States. Emerg. Infect. Dis. 5, 607–625.
Meerburg, B.G., Bonde, M., Brom, F.W.A., Endepols, S., Jensen, A.N.,
Leirs, H., Lodal, J., Singleton, G.R., Pelz, H.J., Rodenburg, T.B.,
Kijlstra, A., 2004. Towards sustainable management of rodents in
organic animal husbandry. Njas-Wageningen J. Life Sci. 52, 195–205.
Meerburg, B.G., Van Riel, J.W., Cornelissen, J.B., Kijlstra, A., Mul,
M.F., 2006. Cats and goat whey associated with Toxoplasma gondii
infection in pigs. Vector Borne Zoonotic Dis. 6, 266–274.
Montoya, J.G., Liesenfeld, O., 2004. Toxoplasmosis. Lancet 363, 1965–
1976.
More, G., Basso, W., Bacigalupe, D., Venturini, M.C., Venturini, L.,
2008. Diagnosis of Sarcocystis cruzi, Neospora caninum, and Toxo-
plasma gondii infections in cattle. Parasitol. Res. 102, 671–675.
Reinken, G., 1998. Production and trade of game and deer meat in
Europe. Zeitschrift Fur Jagdwissenschaft 44, 167–177.
Rhee, M.S., Lee, S.Y., Hillers, V.N., McCurdy, S.M., Kang, D.H., 2003.
Evaluation of consumer-style cooking methods for reduction of
Escherichia coli O157: H7 in ground beef. J. Food Prot. 66, 1030–1034.
Riemann, H.P., Meyer, M.E., Theis, J.H., Kelso, G., Behymer, D.E.,
1975. Toxoplasmosis in an infant fed unpasteurized goat milk. J.
Pediatr. 87, 573–576.
Roberts, T., Frenkel, J.K., 1990. Estimating income losses and other
preventable costs caused by congenital toxoplasmosis in people in the
United-States. J. Am. Vet. Med. Assoc. 196, 249–256.
Roberts, T., Murrell, K.D., Marks, S., 1994. Economic-losses caused by
foodborne parasitic diseases. Parasitol. Today 10, 419–423.
Robson, J.M.B., Wood, R.N., Sullivan, J.J., Nicolaides, N., Lewis, B.R.,
1995. A probable foodborne outbreak of toxoplasmosis. Communi-
cable Dis. Intelligence 19, 517–522.
1370 A. Kijlstra, E. Jongert / International Journal for Parasitology 38 (2008) 1359–1370
Roghmann, M.C., Faulkner, C.T., Lefkowitz, A., Patton, S., Zimmerman,
J., Morris, J.G., 1999. Decreased seroprevalence for Toxoplasma gondii
in seventh day adventists in Maryland. Am. J. Trop. Med. Hyg. 60,
790–792.
Ross, R.D., Stec, L.A., Werner, J.C., Blumenkranz, M.S., Glazer, L.,
Williams, G.A., 2001. Presumed acquired ocular toxoplasmosis in deer
hunters. Retina-J. Retin. Vitreous Dis. 21, 226–229.
Rothova, A., Meenken, C., Buitenhuis, H.J., Brinkman, C.J., Baarsma,
G.S., Boentan, T.N., Dejong, P., Klaassenbroekema, N., Schweitzer,
C.M.C., Timmerman, Z., Devries, J., Zaal, M.J.W., Kijlstra, A.,
1993. Therapy for ocular toxoplasmosis. Am. J. Ophthalmol. 115,
517–523.
Sacks, J.J., Roberto, R.R., Brooks, N.F., 1982. Toxoplasmosis infection
associated with raw goats milk. J. Am. Med. Assoc. 248, 1728–1732.
Schares, G., Vrhovec, A.G., Pantchev, N., Herrmann, D.C., Conraths,
F.J., 2008. Occurrence of Toxoplasma gondii and Hammondia ham-
mondi oocysts in the faeces of cats from Germany and other European
countries. Vet. Parasitol. 152, 34–45.
Schulzig, H.S., Fehlhaber, K., 2006. Seroprevalence of Toxoplasma gondii
in conventionally and organically produced pork and pork-products.
Fleischwirtschaft 86, 106–108.
Scupin, E., 1968. Weitere untersuchungen der überlebensmöglichkeiten von
toxoplasma in schinken. Archiv für Lebensmittelhygiene 12, 11–13.
Sharma, S.P., Baipoledi, E.K., Nyange, J.F.C., Tlagae, L., 2003. Isolation
of Toxoplasma gondii from goats with a history of reproductive
disorders and the prevalence of toxoplasma and chlamydial antibodies.
Onderstepoort J. Vet. Res. 70, 65–68.
Shrestha, J.N.B., Fahmy, M.H., 2005. Breeding goats for meat produc-
tion: a review – 1. Genetic resources, management and breed
evaluation. Small Ruminant. Res. 58, 93–106.
Skinner, L.J., Timperley, A.C., Wightman, D., Chatterton, J.M.W.,
Hoyen, D.O., 1990. Simultaneous diagnosis of toxoplasmosis in goats
and goatowners family. Scand. J. Infect. Dis. 22, 359–361.
Skjerve, E., Waldeland, H., Nesbakken, T., Kapperud, G., 1998. Risk
factors for the presence of antibodies to Toxoplasma gondii in
Norwegian slaughter lambs. Prev. Vet. Med. 35, 219–227.
Sommer, R., Rommel, M., Levetzov, R., 1965. Die überlebensdauer von
toxoplasmazysten in fleisch und fleischzubereitungen. Fleischwirtsch
45, 454–457.
Sroka, J., 2001. Seroepidemiology of toxoplasmosis in the Lublin region.
Ann. Agric. Environ. Med. 8, 25–31.
Sroka, J., Wojcik-Fatla, A., Dutkiewicz, J., 2006. Occurrence of
Toxoplasma gondii in water from wells located on farms. Ann. Agric.
Environ. Med. 13, 169–175.
Stanford, M.R., See, S.E., Jones, L.V., Gilbert, R.E., 2003. Antibiotics for
toxoplasmic retinochoroiditis – an vidence-based systematic review.
Ophthalmology 110, 926–932.
Stanford, M.R., Tan, H.K., Gilbert, R.E., 2006. Toxoplasmic retinoch-
oroiditis presenting in childhood: clinical findings in a UK survey. Br.
J. Ophthalmol. 90, 1464–1467.
Svobodova, V., Knotek, Z., Svoboda, M., 1998. Prevalence of IgG and
IgM antibodies specific to Toxoplasma gondii in cats. Vet. Parasitol.
80, 173–176.
Tassi, P., 2007. Toxoplasma gondii infection in horses. A review.
Parassitologia 49, 7–15.
Tenter, A.M., Heckeroth, A.R., Weiss, L.M., 2000. Toxoplasma gondii:
from animals to humans. Int. J. Parasitol. 30, 1217–1258.
Teutsch, S.M., Juranek, D.D., Sulzer, A., Dubey, J.P., Sikes, R.K., 1979.
Epidemic toxoplasmosis associated with infected cats. N. Engl. J. Med.
300, 695–699.
Thibaut, R., Leproust, S., Chene, G., Gilbert, R., 2007. Effectiveness of
prenatal treatment for congenital toxoplasmosis: a meta-analysis of
individual patients’ data. Lancet 369, 115–122.
Vaillant, V., de Valk, H., Baron, E., Ancelle, T., Colin, P., Delmas, M.C.,
Dufour, B., Pouillot, R., Le Strat, Y., Weinbreck, P., Jougla, E.,
Desenclos, J.C., 2005. Foodborne infections in France. Foodborne
Pathog. Dis. 2, 221–232.
van der Giessen, J., Fonville, M., Bouwknegt, M., Langelaar, M.,
Vollema, A., 2007. Seroprevalence of Trichinella spiralis and Toxo-
plasma gondii in pigs from different housing systems in The Nether-
lands. Vet. Parasitol. 148, 371–374.
Vanknapen, F., Kremers, A.F.T., Franchimont, J.H., Narucka, U., 1995.
Prevalence of antibodies to Toxoplasma gondii in cattle and swine in
the Netherlands – towards an integrated control of livestock produc-
tion. Vet. Q. 17, 87–91.
Vikoren, T., Tharaldsen, J., Fredriksen, E., Handeland, E., 2004.
Prevalence of Toxoplasma gondii antibodies in wild red deer, roe deer,
moose, and reindeer from Norway. Vet. Parasitol. 120, 159–169.
Warnekulasuriya, M.R., Johnson, J.D., Holliman, R.E., 1998. Detection of
Toxoplasma gondii in cured meats. Int. J. Food Microbiol. 45, 211–215.
Weigel, R.M., Dubey, J.P., Siegel, A.M., Kitron, U.D., Mannelli, A.,
Mitchell, M.A., Mateuspinilla, N.E., Thulliez, P., Shen, S.K., Kwok,
O.C.H., Todd, K.S., 1995. Risk-Factors for transmission of Toxo-
plasma gondii on swine farms in Illinois. J. Parasitol. 25, 736–741.
Weinman, D., Chandler, A.H., 1956. Toxoplasmosis in man and swine –
an investigation of the possible relationship. J. Am. Med. Assoc. 161,
229–232.
Woods, M., McDonald, R.A., Harris, S., 2003. Predation of wildlife
by domestic cats Felis catus in Great Britain. Mammal Rev. 33,
174–188.
Zia-Ali, N., Fazaeli, A., Khoramizadeh, M., Ajzenberg, D., Darde, M.,
Keshavarz-Valian, H., 2007. Isolation and molecular characterization
of Toxoplasma gondii strains from different hosts in Iran. Parasitol.
Res. 101, 111–115.