3.Guatteo. Q Fever Impact

Raphal Guatteo, DMV, Msc, PhD, Dipl ECBHM

UMR 1300 Oniris-INRA BioEpAR

Infection

60% asymptomatic

Incubation :2 3 weeks

2% of infected persons

Chronic disease

Endocarditis,

40% Acute Disease- Flu-like syndrom- Hepatitis- Pneumonia- Meningitis

4% need for hospitalization

Pregnancy disorders- Abortions (repeated)- Foetal death- Premature delivery

Adapted from Dupuis, 1985

Brianon (05) 1996

Banon (04) 1987goats

Brasparts (29)2002 goats

Nordheim (67) 2005 goats

Chamonix 2002EwesCholet

2009Veal

Montoison (26)2001 goats

Brianon (05) 1996 Near slaughterhouse Ewes (Eastern day)

Endemic situation around Etang de Berre (13)Annual occurrence +++ > mean occurenceOutdoor Lambing (wind +++)Source : Anses (E. Rousset)

Florac2008 Sheep

foetal serum

Mainly Limited outbreaks but Occurrence similar to

Toxoplasmosis (human aboprtion)

Sometimes outbreak +++

Q fever: a Zoonosis with outbreaks

2009 2357 Human cases 60% asymptomatic 20% pneumonia leading to hospitalization Goats +++ Goats +++

Control actionsBTM MonitoringMandatory vaccination (Phase 1 vaccine)Culling (stamping out) pregnant Goats and/or Ewes

Recent Outbreak in Hungary

Not RareRuminants as main source for human infection

Q fever: Prevalence in Ruminants ?

SpeciesPrevalence at animal levelMedian [Min-Max]

Prevalence at herd levelMedian [Min-Max]

Prevalence at within herd levelMedian [Min-Max]

Cattle 20% [0-100%] 40% [0-100%] 20% [0-60%]

Sheep 11% [0-100%] 25% [0-100%] 30% [0-70%]

Goat 17% [0-100%] 26% [0-100%] 25% [20-85%]

Guatteo et al., 2011

Most frequently Serological study (Risk Assessment ?)

Need for harmonization for Epi study

Human Health IssueDairy Industry Issue

Reduction of Coxiella burnetii shedding Inhalation

RawMilk ?

Inhalation +++

Q fever: Human Health Issue.only?

7

Reduction of Coxiella burnetii shedding

Limit the zoonotic riskSatisfaction of dairy industry

Inhalation

Q fever in Ruminants : Clinical Impact ?

Reproduction: Myth or Reality ?

Adapted from Human pathogenesis Supported by field observations (?)

APSW Abortions and/or then retained placenta, metritis Abortions and/or then retained placenta, metritis Premature delivery Stillbirth Weak Calf syndrome

Other signs: metritis, retained placenta, fertility disorders ? respiratory disorders ?

Invasion and Localisation in the placenta

Establishment of infection

Active infection

Clearance

Immediate clearance of infection

Latent infection

Limited spread Spread to the foetus

Latent infection Active infection WidepreadLocalized

Most Likely outcome: normal offspring

Possible outcomes : APSW syndromeOr normal offspring

Blood spread

Transplacentalspread

LiverAmniotic

fluid

Bowel Lung

Localized or disseminatedinfection

3 complementary approaches

Experimental reproduction Healthy animals Challenged Observed Observed

Case/control study Retrospective or cohort study Difference in prevalence of reproductive disorders depending on

the exposure

Intervention study Implementation of control actions, routine diagnosis Comparison before vs after, with vs without

Experimental Reproduction

Pro Cons

Impact-interaction with other Precise effect of THIS infection

Controlled conditions

Different infectious dose

Impact-interaction with other pathogens

Costly, Limited number of animals

Inoculation often different from natural route

Impact of included animals (egpregnant or not?)

2 studies: one in goat / one in cattle

In goats (Arricau-Bouvery et al., 2005)

0-42 -21 84 100 - 157 6 weeks mating

Vac ph I et 2

0-42 -21 84 100 - 157 6 weeks post abortions

necropsy

Vac ph I et 2 Inoculation

CbC1 104 Not vaccinated group 12 goats Vaccine Phase II group 15 goats Vaccine Phase I group 16 goats

AbortionsDelivery

In goats (Arricau-Bouvery et al., 2005)

Abortions +++Consistent with abortion storm's reported under field conditions

Other signs not investigated

In cattle (Plommet et al., 1973)

12 heifers 8-11 months old Intradermal inoculation Intradermal inoculation 18 months follow-up

In cattle (Plommet et al., 1973)

Abortions +, Subfertility +Consistent with signs reported under field conditions

Lack of detection for other pathogens

Experimental Reproduction: finally

Demonstration Not known

Abortions +++Other reproductive disorders(retained placenta, metritis)

Subfertility +/- Impact of pregnancy stage on susceptibility

Case control studyPro Cons

Retrospective study (cost-effective)Deal with available data

Precise definition of case & controlConcomitant Identification of risk factors

May lead to identify slight effect (size of the dataset)

Precise definition of case & control

If cross sectional survey: association but no causality

May lead to identify statistical effect (size of the dataset) not supported by biology

Case control study in cattle : 7 studies Critical review Exhaustive analysis in the proceeding

Number of animals : > 300 vs < 300: vs o Control group (ie not exposed): vs o Sensitive detection method (ELISA >>FC): vs o

Reference Sample Analysis Reproductive disorders collected/studied

Main results about prevalence

Reliability score

To et al., 207 dairy cows Serum immunofluorescence Metritis, infertility Serum testing: 3.9% PCR

Example of poor design

To et al.,1998

207 dairy cowswho suffered frompreviousreproductivedisorders (93infertility and 114metritis andmastitis

Serum immunofluorescence(IF), serum PCR, milk PCR,milk bacteriology.

Metritis, infertility Serum testing: 3.9% PCR+, milk testing: 24.6%PCR + (Coxiella burnetiihas been isolated in24.6% of PCR + milksamples)

58.9% IF + (phase I)

60.4% IF + (phase II)

O O



Reliability score

Literak and 1397 dairy cows Complement fixation First artificial insemination Correlation between

Example of intermediate quality study

Literak and Kroupa, 1998

1397 dairy cows at dry period (14 herds)

Complement fixation test (phase II antigen)

First artificial insemination (AI1) success rate, final conception rate, calving-AI1 interval, calving interval, average number of AI until pregnancy

Correlation between infection seroprevalence and reproductive performance not statistically significant

o



Reliability score

Lopez-Gatiuset al., 2011

3 herds reproductive disorders (811 dairy cows) (pregnancy rate: 23%,

Serum ELISA Retained placentas, stillbirths, abortions, embryonic resorptions

Correlation between seropositivity and retained placenta

Example of well designed study

(pregnancy rate: 23%, abortion rate: 21%, BTM testing: RT-PCR +, 2 control herds (2371 cows) (pregnancy rate: 38%, abortion rate: 11%, BTM testing:RT-PCR

embryonic resorptions retained placenta (almost twice more of retained placentas in seropositive cows).

No correlation between prevalence and abortion rate after 90 days but low incidence of these events



Reliabilityscore

Ordronneau,2012

2825 cows fromaffected herds (with

Serological test (Cb01strain) performed in

Abortions and retainedplacentas during the 6

Abortion risk increased 2.5times in seropositive

Example of well designed study

abortions) all females older than24 months

following months animals (in comparisonwith seronegative ones)

Risk of retention increased1.5 times in seropositiveanimals (in comparisonwith seronegative ones)

Case control study: finally

Demonstration Not knownDemonstration Not known

Abortions +++

Retained placenta ++

Subfertility +/-

Metritis

Retained placenta itself or retained placenta following abortion

Co-infections ?

Intervention study Field investigation in case of disease: real life Clinical trial (vaccine, antibiotic)

Pro Cons

Concomitant assessment of the effectiveness of control strategies (including placebo)

May lead to hypothesis about the pathogenesis

Comparison of clinically and not clinically affected animals/herds

Inclusion criteria crucial (under or overestimate the treatment efficacy) : diagnosis !!

Measurement of the treatment efficacy itself of the natural course of the infection (ie time)

Costly, limited number of situations (including co-infections)

Field investigation of abortions925 abortions investigated using PCR (Guatteo et al., 2014)

Coxiella burnetii Chlamydophila spp Listeria monocytogenes Salmonella spp. Campylobacter fetus (fetus fetus et fetus venerealis) Anaplasma phagocytophilum BHV4 Leptospira interrogans

Pathogens n %

Coxiella burnetii 160 17,3%Anaplasma phagocytophilum 57 6,2%Salmonella spp 38 4,1%Listeria monocytogenes 16 1,7%Leptospira pathognes 16 1,7%BHV4 15 1,6%Chlamydophila spp 10 1,1%Campylobacter fetus fetus et venerealis 5 0,5%Total 317 34,3%

40

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Cb load

[25-30[[20-25[[15-20[[10-15[

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Placenta Mucus vaginal Avorton Estomac

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Months NQ fever

n %Jan 116 27 23,3%Feb 90 21 23,3%Mar 92 14 15,2%April 84 15 17,9%May 41 11 26,8%May 41 11 26,8%June 48 12 25,0%July 90 15 16,7%Aug 111 22 19,8%Sept 107 6 5,6%Oct 49 7 14,3%Nov 32 3 9,4%Dec 36 4 11,1%

Sample n Fivre Qn %

Aborted calf 30 5 16,7%Stomach content 36 5 13,9%

Placenta 480 89 18,5%Mucus 353 55 15,6%Mucus 353 55 15,6%

PregnancyPregnancyPregnancyPregnancy stage (stage (stage (stage (monthmonthmonthmonth)))) Q Q Q Q feverfeverfeverfever prevalenceprevalenceprevalenceprevalence

[0[0[0[0----1]1]1]1] 0%

]1]1]1]1----2]2]2]2] 20%

]2]2]2]2----3]3]3]3] 6%

]3]3]3]3----4]4]4]4] 14%

]4]4]4]4----5]5]5]5] 19%]4]4]4]4----5]5]5]5] 19%

]5]5]5]5----6]6]6]6] 16%

]6]6]6]6----7]7]7]7] 15%

]7]7]7]7----8]8]8]8] 17%

]8]8]8]8----9]9]9]9] 27%

Days after abortion Weeks after abortion0 1 2 3 1 2 3 4 5 6 7 8+ + + + + + + + - - - -

+ + + + + + + + - - - -

+ + + + + - - - - - -

+ + + + + + + + + - -

+ + + + + + + + + - - -

Goats

+ + + + + + + + + - - -

+ + + + + + + - - - - -

+ + + + + + - - - - -

+ + + + + + - - - - -

+ + + + + + - - - - -

+ + + + + + + - - - -

+ + + + + + + - - - - -

+ + + + + + + - - - - -

Arricau-Bouvery et al. 2005

Cattle

Cow D0 D14 D21 D28PCR result

1 + - - +2 + + + +3 + - - -4 + - - -5 + - - -6 + + + -7 + + - -8 + + - +8 + + - +9 + + + +

10 + + - -11 + - - -12 + - - -13 + + - -14 + - - -15 + - - -16 + - - -17 + - - -18 + - - -19 + - - -20 + - - -21 + - - -22 + - - -23 + + - -24 + - - -

Daprs Guatteo et al. 2012

Studies dealing with vaccine efficacy Both in goats and cattle

In goats mainly focused on abortions mainly focused on abortions experimental conditions

In cattle abortions and fertility 3 studies under field conditions

[[[[ArricauArricauArricauArricau----BouveryBouveryBouveryBouvery et al., 2005, Vaccine]et al., 2005, Vaccine]et al., 2005, Vaccine]et al., 2005, Vaccine]

Vaccination scheme

Vaccine not challenged phase I phase II not vaccinated

Number of goats 27 16 15 12

Duration of gestation (days)ab 150 (+1.8) 153 (+3) 134 (+15) 141 (+8) Abortion (%)b 15 6 87 75 Abortion (%)b 15 6 87 75

Number of kids per goat 1.9 (+0.88) 1.5 (+0.52) 1.67 (+0.62) 1.75 (+0.87) Percentage of goats with contaminated

placenta (%)b ND 37.5 93.3 100

Very strong prevention effect in susceptible animalsLimit : in small ruminants: very high within-herd seroprevalence at abortion time

[Arricau-Bouvery et al., 2005, Vaccine]Vaginal shedding

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NV

Mean duration of sheddingNV 22 days

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DO D1 D2 D3 W1 W2 W3 W4 W5 W6 W7 W8time after kiddings

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NVPh IIPh I

NV 22 daysPh II 16 daysPh I 1,5 day

D-17 D-2 D0 D21 D90 D180 D270 D360CALVING

M, VM, F, BS x 2 M, VM, F M, VM, F M, VM, F M, VM, F M, VM, F

Randomization[Guatteo et al., 2008, Vaccine]

Treatment 1

Vaccination or placebo

Treatment 2

Vaccination or placebo

Determination ofinitial status

Detection of shedding

Shedding follow up among initially susceptible animalsSurvival analysis method (Cox regression model)Description of bacterium load when shedding occurred

6 clinically infected herds336 animals

Initial statusInitial statusInitial statusInitial status

Status Non pregnant Pregnant Total

CowsS 30 62 92

[Guatteo et al., 2008, Vaccine][Guatteo et al., 2008, Vaccine][Guatteo et al., 2008, Vaccine][Guatteo et al., 2008, Vaccine]

40

CowsNS 52 105 157

HeifersS 30 53 83

NS 1 3 4

Total 114 222 336

0.75

1.00

0.75

1.00

0.75

1.00

Efficacy of the vaccine

[Guatteo et al., 2008, Vaccine][Guatteo et al., 2008, Vaccine][Guatteo et al., 2008, Vaccine][Guatteo et al., 2008, Vaccine]

0.00

0.25

0.50

FUP0 100 200 300 400 500

Vaccinated when non-pregnantVaccinated when pregnantPlacebo

Censored animals are marked with circles

0.00

0.25

0.50

FUP0 100 200 300 400 500

0.00

0.25

0.50

FUP0 100 200 300 400 500

Vaccinated when non-pregnantVaccinated when pregnantPlacebo

Censored animals are marked with circles

When vaccinated non pregnant, the risk of becoming shedder was 5 times lower

Vaccination and side effects on reproductionIn France, April of 2005 March of 2006115.562 vaccine phase I (Coxevac) doses: cattle/goat/sheep

- 0.08 % of local reactions

42

- 0.08 % of local reactions - 0.003 % of transitory hyperthermia- 0.002 % of mortality- 0.012 % of abortion- 0.04 % of other (loss of appetite, decrease of milk

production)

Vaccination and side effects on reproductionBoth goats [Arricau-Bouvery et al., 2005] and cattle [Guatteo et al., 2008] studies

Any notable induration Any increase of Body Temperature

DG at D35

Calving between D0 and D35 AbortionEarly calving

DG+ at D35

DG+ at D0Farm

DG at D35

Calving between D0 and D35 AbortionEarly calving

DG+ at D35

DG+ at D0Farm

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at D35

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D0 and D35

11109123total

0012146

0018205

1 (PCR-)023274

0019243

0017172

0120211

at D35at D0

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at D35

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11109123total

0012146

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1 (PCR-)023274

0019243

0017172

0120211

at D35at D0

[Guatteo et al., 2008]

Common treatment for all herds: VACCINATION of Nulliparous

120 Dairy Herds (repeated abortions related to Q fever)

RANDOM

Vaccination Antibiotics NoneVaccinationAntibiotics

In herds with Antibiotics : cows assigned at random: 1 or 2 administrations at drying off 1 administration at calving

Phase 1 Vaccine (Coxevac, CEVA Sant Animale), Long acting Oxytetracycline (20mg/kg, TENALINE LA, CEVA)

M

1 or 2 administrations at drying off and calving

[[[[TaurelTaurelTaurelTaurel et al., 2013]et al., 2013]et al., 2013]et al., 2013]

Comparison of abortions and retained placenta occurrence (-6 : +12 m) between groups

Description of return to service rate between groups

HerdAnimalEnvironment

Vaginal Swabs (22 herds)At calving

BTM or Primiparous mixMonthly or quarterly

Dust, bedding

Effectiveness to reduce/prevent Cb shedding

at individual level Indicator of Cb circulation at herd level

Environmental Cb LoadEffectiveness of bedding removal

1011121314151617

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Within herd abortion rate (median) according to treatment group

3.8

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2.3 2.4 2.1 2.2

0123456789

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Vaccin Vacc + ATBQ ATBQ Control

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before inclusion

after inclusion

Clinical signs Results

Abortions(6 months before abortions vs 12

Vaccination: OR : 0,694 [0.453-1.06] (P=0.09)

Seropositivity (p

Vaccination in nulliparous

Vaccination in Cows ATB at calving

Return to service

(18-26 d)ns ns ns

Improvement of fertility

(18-26 d)Return to service

(27-90 d)OR : 0,538 [0.301-

0.963] (p

Variable N Cb load > 10000 vs. ]0-100] b/mL

OR PVaccination 0.03

Prevention of shedding

None 76 1

After service 73 0.29Before service 13 0.15

Vaccination (cows and heifers) Reduction of Cb load shed when shedding occurred

Reduction of the transmission rate

74 herds, 354 BTM samples, 5254 dairy cows

Vaccination covering rate

0-20%


20-80%


>80%

Reduction of shedding at herd level

Favourable pattern in BTM(reduction of Cbload over time)

0,29 0,17 1

Whole herd vaccination (>80%) : Strong and fast reduction of Cbshedding at herd level

Reduction of shedding in environment (bedding)

80%

100%

Interest of vaccination to reduce Cb loadInterest of bedding renewal to decrease infectious pressure

0%

20%

40%

60%

D0 D90 D180 D270 D360 D450 D540

Negative Low Cb load

Moderate Cb load High Cb load

High ++ Cbload

2 dairy herds PCR positive on bulk tank milk Determination serological status all > 12 months Random allocation (sero status)

301 control and 310 vaccine (Coxevac) at 170 and

[Lopez-Gatius et al., 2014]

301 control and 310 vaccine (Coxevac) at 170 and 190 days of pregnancy

Follow up of return to service, conception rate at first service, calving to conception interval

Results 25 % of cows were seropositive (effectiveness of the

vaccine +++ in seronegative animals)

[Lopez-Gatius et al., 2014]

Vaccine group

Control group

P valuegroup group

Conceptionrate at first service

42% 30% 0,04

Number of AI per pregnancy 1.9 +/-0.1 2.3 +/- 0.2 0,03CCI 92 106 0,02

Intervention study: finally

Demonstration Not known

Abortions +++

Retained placenta ++

Return to service ++

Metritis

Retained placenta itself or retained placenta following abortion

Co-infections ?

To summarize

Species ASPW Complex

Endometritis Fertility disorders

Retained placentaComplex disorders placenta

Cattle + ? + +Goat + ? ? +Sheep + ? ? ?

Outcome Vaccination

Abortions Occurrence

Impaired fertility Return to serviceImpaired fertility Return to service

Coxiella burnetiishedding

Reduction of level of shedding in cows at calvingPrevention of shedding in nulliparous at calving

Whole vaccination : Fast/strong reduction of shedding at cow and herd level

Safety No adverse reactionIncluding in pregnant animals

Acknowledgements

All the cows, farmers and vets included in the studies ....

In memory of Christophe Manteca

Documents

3.Guatteo. Q Fever Impact