Hafez, H.M & R. Hauck

Institute of Poultry DiseasesFree University Berlin

Histomonosis in Germany: Situation since 2004

Governmental regulations

Since 1995there are no products available for treatment

of histomonosis for poultryin the EU.

Governmental regulations

Layer flock Free range

1999

Blackhead disease in layers

(Hafez et al., 2001)

Governmental regulations

Since April 2003 there are no prophylactic

drugs against histomonosis for turkeys available in the EU.

Governmental regulations

Nifursol rest in peace

April 1st 2003

Histomonosis: Current Situation

Date State Affected birds

Age in

Weeks

Total No. Of birds on farm

June 2003 Hessen 9 000 8 9 000

Aug. 2003 Niedersachsen 9 000 5 15 000

Dec. 2003 Baden-Württ. 3 500 7 15 000

April 2004 Niedersachsen 2 500 7 15 000

May 2004 Niedersachsen 2 500 7 25 000

May 2004 Sachsen-Anhalt 4 000 11 8 000

June 2004 Bayern 15 000 8 15 000

Aug. 2004 Niedersachsen 4 500 4 16 000

Sept. 2004 Baden-Württ. 5 000 7 10 000

Feb. 2005 Brandenburg 12 000 5 16 000

Total 10 Outbreaks 66.000 - 144 000

Histomonosis

2005 No. Of samples

No. Of

Positive samples

%

Layers 5 3 60

Broiler breeders 1 1 100

Meat turkeys 19 7

Turkey breeder 6 2 33

Unknown 7 0 0

38 13

Histomonosis

Histomonas

Other Flagellates

Negative-Control

Histomonas

Other Flagellates

Negative-Control

qPCR

Situation in Germany between 2004-2012

0

20

40

60

80

100

120

140

160

180

2004 2005 2006 2007 2008 2009 2010 2011 2012

number of cases number of samples

Number of cases and samples investigatedfor H. meleagridis 2004 - 2012

Cases Samples325 813

0

20

40

60

80

100

120

140

160

pos neg susp

Samples

0

5

10

15

20

25

30

35

2004 2005 2006 2007 2008 2009 2010 2011 2012

pos neg susp

Number of positive and negative cases2004 - 2012

+ - +/-154 169 247% 52% 1%

0

5

10

15

20

25

30

pos neg sus

Seasonal distribution ofpositive and negative samples in %

No. of positive and negative casesin chickens and turkeys

0

2

4

6

8

10

12

14

16

18

2004 2005 2006 2007 2008 2009 2010 2011 2012

Chickens pos Chickens neg Turkeys pos Turkeys neg

0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

100%Chickens

Turkeys

Chickens

Turkeys

Chickens

Turkeys

Chickens

Turkeys

Chickens

Turkeys

Chickens

Turkeys

Chickens

Turkeys

Chickens

Turkeys

Chickens

Turkeys

2004 2005 2006 2007 2008 2009 2010 2011 2012

% of positive and negative casesin chickens and turkeys

No. of positive and negative casesin other species

0

2

4

6

8

10

12

14

2004 2005 2006 2007 2008 2009 2010 2011 2012

Other (mostly peafowl) pos Other (mostly peafowl) neg Not known pos Not known neg

Genotyping of H. meleagridis by C-profiling

- The primers used by van der van den Heijden et al. are designed to amplify also the DNA of other Trichomonads

- As we had samples with trichomonal DNA we designed a primer set more specific forH. meleagridis (Hauck et al. 2010)

Genotyping of H. meleagridis by C-profiling

ssu rRNA 5.8s rRNA lsu rRNAITS 1 ITS 2*Hauck et al, 2010; modified after van der Heijden et al, 2006

1

2

3

4

C-profiling – ITS 1

A

B

C

D

0

5

10

15

20

25

30

35

40

45

Turkey Chickens Peafowl unknownA B C D

C-profiling – ITS 1

Case reports

Turkey breeder farm

Turkey Breeder Farm

1 2 3

2005

Day Cumulative Mortality -House 3

2005 (17W)

0 15 (0.27 %)

1 58 (1.04 %)

2 355 (6.34 %)

3 780 (13.93 %)

4 1437 (25.66 %)

Turkey breeder farm2005

Turkey Breeder Farm

1 2 3

2009

Day Cumulative Mortality - House 2

2009 (8W)

0 6 (0.29 %)

1 40 (1.92 %)

2 60 (2.87 %)

3 114 (5.46 %)

4 298 (14.27 %)

5 754 (36.11 %)

6 1352 (64.75 %)

Turkey breeder farm2009

0

20

40

60

0 1 2 3 4 5 6

cumulative mortality (%)

June 2005,

Week 17 Sept.2009,Week 8

25.7%

(64.8%)

June 2005

September 2009

Type A

C-profling – ITS 1

A

A

- Certified organic production of crops, cattle, pigs, and different species of poultry: Layers, broilers, turkeys, ducks, geese

- Bakery, farm store, holiday apartments

Organic Farm

- Young poultry (all species) is raised for the first 3 weeks in groups of 250 –500 birds in the “garage”

Organic Farm

- Then the groups are moved to different sites:

- “barn” with 5 compartments on 3 floors- “duck house”, 1 compartment- “pig house”, 1 compartment- layer house, 3 x 1400 layers- 6 mobile barns

Organic Farm

August 2005Mobile barnType BLayer flock:H. gallinarumNo Histomonas

8-9 week

August 2005Mobile barnType BLayer flock:H. gallinarumNo Histomonas

May 2008Mobile barnTyp A

8-9 week

August 2005Mobile barnType BLayer flock:H. gallinarumNo Histomons

May 2008Mobile barnTyp A

Fall 2008

„Pig house “Type BBroiler

„Barn“Type A Turkeys

„Barn“

„Duck house“Broiler Typ ?

Layer flock (seperate barn):No Histomonas

8-9 week

55 days

9 weeks

60 days

0

100

200

300

400

500

October November December

Mortality in Broilers

Total: 67 %

Total: 100 %

- High mortality in broilers certainly not (only) the result of the infection with H. meleagridis

- Co-infections of H. meleagridis and E. tenella are known to aggravate symptoms* *Hu & McDougald, 2001

Organic Farm

- All typed strains were either type A (turkeys May 2008 and fall 2008) or B (turkeys August 2005 and broilers fall 2008)

- We were not able to show, that the chickens were the source of infections for the turkeys

- Persistence of the causing strains on the farm is possible

Organic Farm

Week 4-5

Move to other barn100

200

300

400

500

600

700

53 54 55 56

cum

ulat

ive m

orta

liti

es

1 4 1525

57 58 59 60

54

152

183

238

Turkey-Farm

House 1 House 2

Wire mesh

• Separate entrances to the toms on the other side of the houses with disinfection

• Second wire mesh with 1 m distance• Addition of new litter every day• Natural products via drinking water immediate (Ropadiar)

• Natural products via feed next day

no effect

Turkey-Farm

Immediate measures

Turkey-Farm

• After toms were euthanized:– Litter from house 2

was brought to house 1– Backpart of house 2 was

cleaned and disinfected– Fresh litter – Hens from house 1 were brought to house 2

• House 1 with litter – house 2 with hens

2620 hens

2620 hens

House 1 House 2

Control of litter

• Litter taken after the euthanization for control using PCR Result

Negative

Litter taken after the euthanization for Litter taken after the euthanization for parasitologyparasitology

Result

Negative

• Cloacal swabs were taken and contolled usingPCR

Date PCR result

10.02.2007 Negative

21.03.2007 Negative

Control of hens

Neither symptoms nor mortality could be observedin the hens all the time.The hens were slaughtered at week 16As well as the toms between these hens

Attempted treatments with “alternative” products in field cases of histomonosis in turkeys

Product Ingredients (manufacturer) Application

AD3EC Vitamins A, D3, E, and C 0.05% in water

Biostrong Microencapsulated essential oils, herbal

compounds, and an acid complex

0.05% in feed

Hemmopain Vitamin E, methionin, and herbs 0.05% in water

Histosan Herbal extracts and essential oils 0.3% in feed

Klat-Arom Flavouring substances 0.01% in water

Mineral flüssig Various minerals 0.05% in water

Ropadiar Oregano extract 0.02% in water

Spuracid Various trace elements and minerals,

especially copper

0.2% in water

Ventrarctin Extracts of chamomile, peppermint, common

yarrow, and sorbic acid

6% in water

Histomonosis – Control approaches

Killing

Slaughter

or

Histomonosis – Control approaches

Killing

Histomonosis – Control approaches

Killing

Histomonosis – Control approaches

0

1

2

3

4

4 5 7 8 11

Age in weeks

Histomonosis – Control approaches

Killing

Histomonosis – Control approaches

*- CO2 - Gas in container

*- CO2 - Gas directly in poultry house

*- Electric water bath system

*- Neck dislocation

*- T61- injection

*- CO - Gas in small container

*- CO - Gas directly in poultry house

Stamping out

Histomonosis – Control approaches

Histomonosis – Control approaches

Histomonosis – Control approaches

CO2 - Gas in container

Histomonosis – Control approaches

Mobile electrical water bath system

Histomonosis – Control approaches

neck dislocation

Histomonosis – Control approaches

T 61 injection

Histomonosis – Control approaches

Rendering

Disposal of carcasses

Histomonosis – Control approaches

Dr. Christina PoppPoultry Health Service, Stuttgart, Baden-Württemberg

Dr. Johannes AkaMoorgut Kartzfehn von Kameke, Bösel, Germany

Stefanie BalczulatInstitute of Poultry Diseases, Free University, Berlin

Acknowledgment

Future approaches

Thank you !

Treatment trials Active substance Therapy Effect

Ropadiar sol 7,5%

(Water)

Oreganum Extract negative

Ropadiar

(Feed)

Oreganum Extract negative

Protophyt

(Water)

Chinese Cinnamon,

Cinnamon, Garlic

negative

Valbacen

(Water)

negative

(Redmann et al. 2003)

Effect of several products on H. meleagridis in infected flocks

Changing the litter directly after

the onset of clinical signs was

not effective in reducing the mortality

Blackhead – Control approaches

pos neg susp2004 12 212005 13 312006 10 142007 16 112008 14 142009 21 142010 24 27 22011 31 232012 13 14Total 154 169 2

Number of positive and negative cases2004 - 2012

total pos neg suspCaecum 246 115 130 1Liver 228 91 134 3Caecum + liver pooled 62 30 31 1Other organs/cloacal swabs/feces

63 15 48 0

Invertebrates 67 16 49 2Environment/feed 117 45 71 1Others 7 1 6 0

790 313 469 8

Samples

Invertebraten