Jacques Guillot, Guillaume Le Loc’h, Pascal Arné, Françoise Féménia, René Chermette - PowerPoint PPT Presentation

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Jacques Guillot, Guillaume Le Loc’h, Pascal Arné, Françoise Féménia, René Chermette. Avian aspergillosis. UMR INRA, AFSSA, ENVA, UPVM 956, Biologie Moléculaire et Immunologie Parasitaires et Fongiques, Ecole Nationale Vétérinaire d’Alfort, 94704 Maisons-Alfort Cedex, France. Introduction.

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Jacques Guillot, Guillaume Le Loc’h, Pascal Arné,

Françoise Féménia, René Chermette

Avian

aspergillosis

UMR INRA, AFSSA, ENVA, UPVM 956, Biologie Moléculaire et Immunologie Parasitaires et Fongiques, Ecole Nationale Vétérinaire d’Alfort, 94704 Maisons-Alfort Cedex, France


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Introduction

Aspergillosis is considered as one of the most important infectious diseases in birds

• early 1800s: first observations

scaup duck, jay, bullfinch, bustard and several swans

• 1898: first cases in turkey poults (Lignières & Petit in France)

• Since then, cases in a very wide range of avian species…


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5 key questions…

Why are birds more susceptible than mammals ?

In which circumstances does aspergillosis occur in birds ?

What do the main clinical signs and lesions look like ?

Is it possible to make an early diagnosis in birds ?

Is it possible to treat or prevent avian aspergillosis ?


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Susceptibility of birds

Birds are much more susceptible to aspergillosis than mammalian species

• Environmental contamination by Aspergillus spp.


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Susceptibility of birds

lungs / air sacs

Birds are much more susceptible to aspergillosis than mammalian species

• Environmental contamination by Aspergillus spp.

• Avian anatomy and physiology

40-43°C


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Susceptibility of birds

Birds are much more susceptible to aspergillosis than mammalian species

• Environmental contamination by Aspergillus spp.

• Avian anatomy and physiology

• Avian immunology

Lack of resident macrophages in airways and air sacs

Heterophils without peroxydation


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Epidemiology

Aspergillus fumigatus / Aspergillus flavus, Aspergillus niger…

humidity, dampness

drying period …

Overgrowth

Specific virulence factors ?


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Epidemiology

An epidemiological survey was conducted in France

• a 600 m2 confinement building in the Center of France

• a flock comprising 4500 turkeys

females slaughtered at the age of 12 w and males at the age of 16 w

Genotyping of environmental and clinical A. fumigatus isolates

(2 microsatellite markers)

Bart-Delabesse et al. J. Clin. Microbiol. 1998


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Epidemiology

Lair-Fulleringer et al. Poultry Science 2006


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Epidemiology

5 females

5 males

10 healthy chicks,

23 isolates,

1 unique genotype

Lair-Fulleringer et al. J. Clin. Microbiol. 2003


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Epidemiology

9 healthy turkeys, 55 isolates, 17 genotypes

Lair-Fulleringer et al. J. Clin. Microbiol. 2003


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Epidemiology

2 carcass condemnations, 36 isolates,

2 genotypes

Lair-Fulleringer et al. J. Clin. Microbiol. 2003


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Epidemiology

362 air samples, 134 isolates

53 genotypes

16 week-sampling period

Lair-Fulleringer et al. Poultry Science 2006


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Epidemiology

Aspergillus fumigatus / Aspergillus flavus, Aspergillus niger…

Overgrowth

Specific virulence factors ?

captive environment

handling

migration…

• Stress

• Other micro-organisms

• Toxicosis

• Therapeutics

turkeys, quails, raptors, penguins, parrots, waterfowl

• Species

• Avian strains

• Individual susceptibility ?

• Age

• Sex

mycotoxins ?

corticosteroids


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Clinical signs and lesions

Different avian species

Different epidemiological situations

Many clinical signs

Acute aspergillosis

= inappetance, depression, polydipsia, polyuria, dyspnoea, cyanosis

= sometimes, sudden death without any signs

fatal evolution

Chronic aspergillosis

= signs are dependent on the area of invasion

change in voice

respiratory stridor

exercice intolerance

ataxia, torticollis, seizures

conjunctivitis, keratitis

beak malformation…


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Clinical signs and lesions

Aspergillosis

of eggs

air sacculitis

Kaminski et al.

ENVA

ENVA


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Clinical signs and lesions

meningo-

encephalitis

pneumonia

Séguin

Chute

arthritis

uveitis

Séguin

Séguin


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Diagnosis

Radiology

Endoscopy

Hematology

Biochemistry

Electrophoresis

Cytology, histology

Mycological culture

Serology

PCR ?

Falcons, parrots, wild birds in zoological or rehabilitation centers

Clinical signs

Epidemiology


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Diagnosis

Falcons, parrots, wild birds in zoological or rehabilitation centers

Clinical signs

Epidemiology

Radiology

Endoscopy

Hematology

Juliet Joseph,

Abu Dhabi Falcon Research Hospital

Biochemistry

Electrophoresis

Cytology, histology

Mycological culture

Serology

PCR ?


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Diagnosis

Falcons, parrots, wild birds in zoological or rehabilitation centers

Clinical signs

Epidemiology

Radiology

Endoscopy

Hematology

Biochemistry

Electrophoresis

Cytology, histology

Mycological culture

Serology

a promising approach ?

PCR ?


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Diagnosis

Serology

• Immunologically, birds respond to Aspergillus infection in the same way as mammals and a type I response appears most beneficial

• Birds also respond with specific antibody production similar in its kinetics to mammals

• Serological tests that may be used in birds include the detection of specific antibodies or fungal antigens


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Diagnosis

Serology

 detection of specific antibodies

High prevalence of seropositivity in captive penguins

Low prevalence of seropositivity in wild birds

False negative results

some birds may not be able to mount an appropriate response

some infection locations result in limited antigenic stimulation

Antibody titers not necessary correlated to clinical severity


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Diagnosis

Serology

 detection of galactomannan

• agglutination (Pastorex Aspergillus®)

• polyclonal sandwich ELISA

• monoclonal sandwich ELISA (Platelia Aspergillus®)


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Diagnosis

Serology

 detection of galactomannan

839 serum samples (from suspected or confirmed cases)

Galactomannan in 50% of samples from penguins

in 25% of samples from other birds

Many chronic cases = negative for antibody but positive for galactomannan

But long term chronic cases = negative for both antigen and antibody !

Cray et al. ISHAM 2006 (poster P-0011)


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Diagnosis

Serology

 detection of galactomannan

90 serum samples (from cases in falcons)

182 control serum samples (from healthy falcons)

Galactomannan in 12% of samples from infected falcons

in 5% of samples from healthy birds

Arca-Ruiba et al. Vet. Rec. 2006


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Diagnosis

Serology

 detection of galactomannan

Le Loch’ et al. ISHAM 2006 (poster))


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Diagnosis

Serology

 detection of galactomannan

Many false negative results

variable release of GM

variable kinetics

according to avian species

according to physiological status of each bird

according to the level of immune complexing

Some false positive results

circulating GM from other fungi

cross reactivity with bacterial components (food supplementation ?)


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Diagnosis

Falcons, parrots, wild birds in zoological or rehabilitation centers

Clinical signs

Epidemiology

Radiology

Endoscopy

Hematology

Biochemistry

Electrophoresis

Cytology, histology

Mycological culture

Serology

Hardy et al. AAV proc. 2003

Dahlahausen et al. AAV proc. 2004

PCR ?


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Diagnosis

Radiology

Endoscopy

Hematology

Biochemistry

Electrophoresis

Cytology, histology

Mycological culture

Serology

Necropsy

Turkeys, chickens, quails, ducks, ostriches…

Clinical signs

Epidemiology


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Treatment

Falcons, parrots, wild birds in zoological or rehabilitation centers

Amphothericin B

Flucytosine

Ketoconazole

Itraconazole

Terbinafine

Voriconazole (Langhofer, AAV proc. 2004)

Radiography, endoscopy or serology for the follow up

+ Supportive therapy

+ Surgical debridement of the lesions


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Treatment

Turkeys, chickens, quails, ducks, ostriches…

No treatment !


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Prevention

• Reduction of fungal contamination

• Reduction of stress

• Chimioprevention ?

• Vaccination

Richard (1984) reduced mortalities by 50% in turkey poults vaccinated with germinated A. fumigatus conidia

Vaccination with a heat-killed culture filtrate preparation has been reported to reduce mortality in ducks and waterfowl


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Conclusions

« Avian aspergillosis »

= not a single entity but a complex of several diseases…

• need for specific avian models of aspergillosis

• variable epidemiological situations

• several diagnostic tools required

• poor prognosis

• importance of prophylactic procedures


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References

  • Cray, C., Rodriguez, M. & Watson, T. (2006) Aspergillus serodiagnostics in avian species. 16th Congress of the International Society for Human and Animal Mycology (ISHAM), June 25-29th 2006, Paris (Poster).

  • Harmon, B. (1998). Avian heterophils in inflammation and disease resistance. Poultry Science, 77, 972-977.

  • Kearns, K.L. (2003). Avian aspergillosis. In: Recent advances in avian infectious diseases. Kearns KS, Loudis B (Eds). Ithaca, International Information Service.

  • Klika, E., Scheuermann, D.W., De Groodt-Lasseel, M.H.A., Bazantova, I. & Switka, A. (1996). Pulmonary macrophages in birds (barn owl, Tyto tyto alba), domestic fowl (Gallus gallus domestica), quail (Coturnix coturnix) and pigeon (Columbia livia). Anatomy Record, 246, 87-97.

  • Kunkle, R.A. & Rimler, R.B. (1996). Pathology of acute aspergillosis in turkeys. Avian Diseases, 40, 875-886.

  • Lair-Fulleringer, S., Guillot, J., Desterque, C., Seguin, D., Warin, S., Chermette, R. & Bretagne, S. 2003. Differentiation of Aspergillus fumigatus isolates from breeding turkeys and their environment by genotyping with microsatellite markers. Journal of Clinical Microbiology, 41, 1798-1800.

  • Le Loc’h, G., Arné, P., Bougerol, C., Risi, E., Péricard, J.M., Quinton, J.F., Bretagne, S. & Guillot, J. (2006) Detection of circulating serum galactomannan for the diagnosis of avian aspergillosis. 16th Congress of the International Society for Human and Animal Mycology (ISHAM), June 25-29th 2006, Paris (Poster).

  • Morris, M.P. & Fletcher, O.J. (1988). Disease prevalence in Georgia turkey flocks in 1986. Avian Diseases, 32, 404-406.

  • Peden, W.M. & Rhoades, K.R. (1992). Pathogenicity differences of multiple isolates of Aspergillus fumigatus in turkeys. Avian Diseases, 36, 537-542.

  • Redig, P.T., Post, G.S., Concannon, T.M. & Dunette, J. (1986). Development of an ELISA for the detection of aspergillosis in avian species. Proceedings of the Association Avian Veterinarians, 165-178.

  • Redig, P.T. (1993). Avian aspergillosis. In: Fowler ME (Ed) Zoo and wild animals medicine. WB. Saunders Company, Philadelphia, 178-181.

  • Richard, J.L. (1997). Aspergillosis. In: Diseases of poultry. Calmek B.W. (Ed), Mosby-Wolfe, London, 351-365.

  • Taylor, J.J. & Burroughs, E.J. (1973). Experimental avian aspergillosis. Mycopathologia Mycologia Applicata, 51, 131-141.

  • Tell, L.A. (2005). Aspergillosis in mammals and birds: impact in veterinary medicine. Medical Mycology, 43, S71-S73.


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