Toxoplasma and food - old.iss.itold.iss.it/binary/crlp/cont/TOXOPLASMOSIS_BUXTON.1190113883.pdf ·...
Transcript of Toxoplasma and food - old.iss.itold.iss.it/binary/crlp/cont/TOXOPLASMOSIS_BUXTON.1190113883.pdf ·...
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Toxoplasmosis is arguably….. “the most widespread and prevalent protozoan parasitic zoonosis”.
(Macpherson 2005)
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Toxoplasma and food1 Toxoplasma gondii
parasite / animals /environment2 Zoonosis
infection vs disease3 Control
detection / reduction / education
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4,000 million years ago life on earth begins 1,500 mya Protozoa appear1,500-500 mya Apicomplexan protozoa develop
major clades todaypiroplasms (Theileria, Babesia)haemosporinids (Plasmodium)gregarinescryptosporidiansenteric coccidia (Eimeria, Cyclospora)tissue cyst forming coccidia (Toxoplasma, Neospora,
Sarcocystis)LD Sibley Vet Parasitol 2003
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tissue cyst forming coccidia(Toxoplasma, Neospora, Sarcocystis, Hammondia, Besnoitia)
common ancestor - 250 myamammalian hosts diverge and expand over similar periodLD Sibley Vet Parasitol 2003
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gundi (Ctenodactylus gundi)1908 Nicolle ManceauxPasteur Institute, Tunis
rabbit (Oryctolagus cuniculi) Splendore 1908 Sao Paulo Brazil
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Toxoplasma gondiisubphyllum Apicomplexafamily Sarcocystidaeobligate intracellular protozoan toxon (arc) plasma (form) Greek
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Toxoplasma gondiiAsexual cycle
BRADYZOITESin tissue cysts
TACHYZOITES
all warm-blooded animals
Likely mode of transmission Toxoplasma cysts or an “as yet unrecognised”form of the parasite (Hartley 1966)
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Toxoplasma gondiiAsexual cycle
BRADYZOITESin tissue cysts
TACHYZOITES
all warm-blooded animals
Hutchison 1965 Hutchison 1965 (+ many others 1970)(+ many others 1970)
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Toxoplasma gondiiAsexual cycle Sexual cycle
BRADYZOITESin tissue cysts
Gut infection
OOCYSTSTACHYZOITES
all warm-blooded animals
cats
ingestion
ingestion
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Cats and Toxoplasmaoocyst excretion
ingestion day oocyst excretion
oocysts 20 50% thousandstachyzoites 20 50% thousandsbradyzoites 4-10 100% millions
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Toxoplasma gondii isolates very similarFerguson Hutchison (1981) Ann Trop Med Parasitol
Molecular strain typing(Restriction Fragment Length Polymorphisms – RFLP)
>95% discrete clonal groupsType I acutely virulent to miceType II non-virulent to miceType III non-virulent to mice
Sibley Boothroyd (1992) Nature
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0
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Type I Type II Type III
AIDSCongenAnimal
Toxoplasma gondiiTypes I, II and III
Howe and Sibley (1995) J Inf Dis
4 isolates mixed lineage
percent(n = 106)
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Type I Type II Type III
Clin Toxo
Toxoplasma gondiiTypes I, II and III
clinical human cases
Howe Honore Derouin Sibley (1997) J Clin Micro
percent(n = 68)
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Toxoplasma gondii
Type II clinical human toxoplasmosispersistent animal infections
high prevalence pigs and sheepHowe Honore Derouin Sibley (1997) J Clin Micro
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Toxoplasma in animals
Clinical toxoplasmosis =T. gondii LIVE in environmentBUT tip of the iceberg
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Toxoplasma in animals
Clinical toxoplasmosis =T. gondii LIVE in environmentBUT tip of the iceberg
Serology infection WIDESPREAD
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Toxoplasma in animalslandwater
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Ovine toxoplasmosis - temperate sheep rearing countriesNew Zealand, Norway, UK, Eire, Spain, Portugal, France
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minor route
persistently infectedEWES
infected LAMB
abortion
WILD MICE
primary infection
tissue cysts
oocysts
contam.food/water
transmission of ovine toxoplasmosis
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oocysts may contaminate grass
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Toxoplasmosis and Risk factorsSeroprevalence of Toxoplasma sheep France(slaughterhouse survey/clin normal) (Dumètre et al 2006)
Lambs (n=164) 22% sero+ve
Ewes (n=93) 65.6% sero+ve
Type II Tg (n=50) 100%
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oocysts may also contaminate grain
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Toxoplasma oocyst contamination of livestock feed
Oocysts in cat faeces 1 million/g50g cat faeces in 10 tonnes feed5,000 oocysts /kg feed100 oocysts will infect one ewe50 sheep infective doses/kg feed
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50 infective doses/kg feed
oocysts in hay
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Pigs become infected when reared outsideKijlstra et al 2004
Reindeer seroconvert (age related) Oksanen et al 1997
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Buxton et al 1997
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T. gondii in game/wildlife (worldwide)% n
bear 15-84 (1,828)wild boar 15-37 (1,131)deer 7-60 (3,017)moose 15 (125)reindeer 0-25 (2,577)gazelle/antelope 0-37 (694)(Tenter et al 2000)
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Toxoplasma in the environmentlandwater
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Toxoplasma and aquatic wildlife
Wild otters - T. gondii
Sea otters (Enhydra lutris nereis) Californian coast16% mortality Toxoplasma gondii infection (n = 35)Type II (40%) and Type x (60%) isolatedType x associated with meningoencephalitis
Miller et al (2004)
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Toxoplasma and aquatic wildlife
Wild otters - serology for T. gondii
River otters (Lutra canadensis) North Carolina45% +ve (46/103) Tocidlowski et al (1997)
Sea otters (Enhydra lutris nereis) Californian coast52% +ve (115/223) Miller et al (2002)
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Beluga whale (Delphinapterus leucas)Toxo (n = 2) 6/22 sero+ve St Lawrence estuary Mikaelian et al (2000)
Spinner dolphin (Stenella longirostris)Disseminated Toxo Hawai Migaki et al (1990)
Striped dolphin (Stenella coeruleoalba)Toxo (+/- morbillivirus) (n = 4) Med. Domingo/Agrimi et al (1995)
Risso’s dolphin (Grampus griseus)Fatal maternal/foetal inf. Med. Resendes et al (2002)
Minke whale (Balaenoptera acutorostrata)100% sero-ve (n = 202) Nth Atlantic/Arctic Ocean
Oksanen et al (1998)
Toxoplasma and sea mammals(cetaceans)
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Harbor (Common) seal (Phoca vitulina)Toxo + S neurona California Miller et al (2001)
Harbor (Common) seal (Phoca vitulina)7.6% sero+ve (29/380) Puget Sound Washington
Lambourn et al (2001)Harp seal (Phoca groenlandica)Ring seal (Phoca hispida)Hooded seal (Cystophora cristata)
100% sero-ve (n = 443) Nth Atlantic/Arctic OceanOksanen et al (1998)
Toxoplasma and sea mammals(pinnipeds)
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Water Potential source of zoonotic protozoal infectionsGiardia sppCryptosporidium sppToxoplasma gondii
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Toxoplasma in the environmentLandInfection (+/- disease) widespreadfarm livestock and wildlifeWaterInfection (+/- disease)Associated urban areasbut not always –Vancouver Island Canada 1994/1995
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Toxoplasma and food1 Toxoplasma gondii
parasite / animals / environment2 Zoonosis
infection vs disease3 Control
detection / reduction / education
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infection
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Human infection Seroprevalence surveys
North America, Europe 32% (n=17 1956-97)Central/Sth America, Caribbean 38% (n=17 1956-96)African Countries 42% (n=18 1977-96)Middle East, Sth Asia, Pacific 30% (n=27 1973-97)
(Hall Ryan and Buxton 2001)
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2
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4535
60Stray-Pedersen
Toxoplasma antibodies20 year olds/100
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Kodym et al (2001) Intl J Parasitol, 31, 125-132
Seroprevalence of toxoplasmosis in the Czech Republic
% +ve
Age (years)
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Seroprevalence of toxoplasmosis in the Czech Republic
Kodym et al (2001) Intl J Parasitol, 31, 125-132
% +ve
Age (years)
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Seroprevalence of toxoplasmosis in the Czech Republic
Kodym et al (2001) Intl J Parasitol, 31, 125-132
% +ve
Age (years)
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Human toxoplasmosis
Infection vs Disease
Infection commonup to 80%
Disease rareConfined to risk groups?
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Human toxoplasmosis
Risk groupsImmunosuppressed - acute (fatal) illness
AIDScancer therapycertain cancers (lymphoma)transplant recipients (+ infected transplant)
Pregnancy - congenital infection
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Human toxoplasmosis
Acute illnessfever headache listlessness muscle acheslymphadenopathy (enlarged tender lymph nodes)
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chorioretinitis(pre & post natal)
pneumonia (interstitial)encephalitis
hepatitismyositis
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Congenital toxoplasmosis
60% mothers no symptoms40% vague symptomsfever headache listlessness muscle acheslymphadenopathy
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1
1-2 31-3 3
3-62-3
2
Congenital Toxoplasma infection per 1000pregnancies
Stray-Pedersen
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Congenital human toxoplasmosisprimary infection initiated during pregnancy
1st trimester infection 15% fetuses outcome “severe”
2nd trimester infection 30% fetuses outcome “mod”
3rd trimester infection 60% fetuses outcome “mild”Desmonts &Couvreur 1984
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Toxoplasma and pregnancy
gestation1st 2nd 3rd trimester
Risk of placental transmission
Risk of fetal damage
+
+++
++
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Congenital human toxoplasmosis
fetal death CNS lesions - hydrocephalus spasticitymild clinically inapparent
longterm 1 in 3 require medical caremental changes/retardation/learning difficultiesvisual handicaps (chorioretinitis)
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BUT clinical toxoplasmosisand link to food
NOT ONLY in risk groups
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Human toxoplasmosisoutbreaks linked to food3 of 4 men (Korea) unilateral chorioretinitis“feasted” on raw pork, liver, spleen of wild pig “in a belief of its special nutritional value”Patient 1 recovered with treatment (retinal scarring)Patients 2 and 3 remained blind despite treatment
Won-Young et al (1997) J Inf Dis 175 1280-1282
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Human toxoplasmosisoutbreaks linked to food
5 of 11 soldiers lymphadenopathyate raw pork, liver of domestic pig
Sulphonomide/pyrimethamine 1 monthFull recovery
Won-Young et al (1997) J Inf Dis 175 1280-1282
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Human toxoplasmosisOutbreak linked to drinking water Canada Vancouver Island 1994/95
Small reservoir supplying unfiltered chlorinated water to 292,000 people
100 cases acute toxoplasmosis3 times expected incidence
Bowie et al (1997) Lancet 350 173-177
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Human toxoplasmosisOutbreak linked to drinking water Canada Vancouver Island 1994/95100 cases acute toxoplasmosis
51 lymphadenopathy19 retinitis11 malais/fever/headache18 no symptoms1 no history
(includes pregnant/post natal women out of 3182 screened)Bowie et al (1997) Lancet 350 173-177
Toxoplasma infection in local feral cats cougars (oocysts in faeces x1)
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Toxoplasmosis and Risk factorsmost Toxoplasma/ocular disease POSTNATAL
retinochoroiditis
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Toxoplasmosis and Risk factorsmost Toxoplasma/ocular disease POSTNATAL
(questionaire - Jones et al 2006)Increased risk raw/lightly cooked meat
cured/dried/smoked meateating “frozen lamb”garden/yard
Decreased risk raw beef/ground beefraw chicken/ground chicken
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Toxoplasmosis and Risk factorsPork and Toxoplasma Brazil
Finishing pigs 17% sero+ve (n=286)(de A Dos Santos et al 2005)
Pork sausages (fresh) live Toxoplasma 8.7%(Dias et al 2005) (n=149 8 factories)
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sources of infectionoocysts
cat litter traysgarden – children’s sandpits unwashed fruit / vegetablescontaminated water
tissue cysts / tachyzoitesraw/lightly cooked meataborting sheep (goats pigs)
Human toxoplasmosis
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Toxoplasma and food1 Toxoplasma gondii
parasite / animals / environment2 Zoonosis
infection vs disease3 Control
detection / reduction / education
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DETECTION of DETECTION of T. gondiiT. gondiiclinical signsclinical signsserology IFAT ELISA LATserology IFAT ELISA LAThistopathologyhistopathologyPCR (B1 & P30/SAG genes)PCR (B1 & P30/SAG genes)inoculation (mice TC)inoculation (mice TC)
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Killing Toxoplasma
T. gondii tissue cystssurvive up to 3 min 640C in pork
(Dubey et al 1990)Cook red meat until brown
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Killing Toxoplasma
High Pressure Processing (HPP)Non thermal treatment
extends shelf-liferetains flavour colour vitamins
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Killing Toxoplasma
High Pressure Processing (HPP)Non thermal treatment
extends shelf-liferetains flavour colour vitamins
T. gondii tissue cysts (in ground pork)Survive 0, 100, 200 MPa for 90 secondsKilled by 300MPa for 30 seconds(Lindsay et al 2006)
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Killing Toxoplasma oocysts
formalin 10% 24hammonia 5.5% 1hethanol 95% 1h
20% 7diodine (tinct) 7% 10m
2% 3hH2O 500C 30mH2O 20/220C 548dH2O 40C (2%H2SO4) 548dDubey and Beattie 1988
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Control of toxoplasmosis in animalsmanagement
herbivore - omnivore - carnivoreavoid contamination food/water - oocysts
omnivore - carnivoretreat infected meat (tissue cysts - cooking)
vaccination ovine toxoplasmosis - live vaccine (Toxovax)
pharmaceuticals
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Toxovax(Intervet BV)
Ovine toxoplasmosis control - vaccination
live toxoplasma vaccine (S48 tachyzoite suspension)live toxoplasma vaccine (S48 tachyzoite suspension)
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Toxovax (S48 T. gondii) and sheep
Will vaccination control clinical disease?YES
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Toxovax (S48 T. gondii) and sheep
Will vaccination produce tissue cysts?NO S48 T. gondii “incomplete”
cleared within 3 weeks
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Toxovax (S48 T. gondii) and sheep
Is meat from vaccinated sheep safe to eat?YES 4 weeks post vacc. no S48 T. gondii in meatBUT “wild” strain T. gondii may be in meat
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Toxovax (S48 T. gondii) and sheep
If vaccinated sheep ingest T. gondii oocystswill they develop tissue cysts? S48 suppresses/prevents multiplication of ToxoplasmaS48 suppresses/prevents spread in lymph and blood
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Toxovax (S48 T. gondii) and sheep
Can vaccination control/reduce numbers T. gondii tissue cysts in muscle (meat)?
Answeryes
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ControlAttempt to reduce toxoplasma oocyst contamination of water (assuming infection is derived from water run-off from the land)Captive animals possible?Wild animals difficult
Toxoplasma and sea mammals
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pharmaceuticals
toxoplasmosis
● sulphonamides● pyrimethamine● trimethoprim / sulphadiazine● baquiloprim / sulphadimidine● clindamycin
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Toxoplasma and food1 Toxoplasma gondii
parasite / animals / environment2 Zoonosis
infection vs disease3 Control
detection / reduction / education
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0102030405060708090
100
1965
1966
1969
1973
1987
1990
1995
ParisLiegeStockholmSth York
Toxoplasma IgG in pregnant women
Gilbert 2000