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

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…

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 ?

Susceptibility of birds

Birds are much more susceptible to aspergillosis than

mammalian species

• Environmental contamination by Aspergillus spp.

 Susceptibility of birds

Birds are much more susceptible to aspergillosis than

mammalian species

• Environmental contamination by Aspergillus spp.

• Avian anatomy and physiology

lungs / air sacs

40-43°C
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

Epidemiology

Aspergillus fumigatus / Aspergillus flavus, Aspergillus niger…

humidity, dampness
Overgrowth drying period …

Specific virulence factors ?

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
Epidemiology

150
125
100
75
50
25
0
3 4 5 6 7 8 9 10 11 12 13 14 15 16

Aspergillus flavus Aspergillus fumigatus Other fungi

Lair-Fulleringer et al. Poultry Science 2006

Epidemiology

5 males 5 females

10 healthy chicks,
23 isolates,
1 unique genotype

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

Epidemiology

9 healthy turkeys, 55 isolates, 17 genotypes

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

Epidemiology

2 carcass condemnations,
36 isolates,
2 genotypes

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

Epidemiology

362 air samples, 134 isolates

53 genotypes

16 week-sampling period
Lair-Fulleringer et al. Poultry Science 2006
Epidemiology

Aspergillus fumigatus / Aspergillus flavus, Aspergillus niger…

Overgrowth
Specific virulence factors ?

captive environment
turkeys, quails, • Stress handling
• Species raptors, penguins,
migration…
parrots, waterfowl
• Avian strains • Other micro-organisms
• Individual susceptibility ?
• Age • Toxicosis mycotoxins ?
• Sex • Therapeutics corticosteroids
 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…
Clinical signs and lesions

Aspergillosis
of eggs

air sacculitis

ENVA

Kaminski et al.

ENVA
Clinical signs and lesions

meningo-
encephalitis
pneumonia

Séguin
Chute

uveitis arthritis

Séguin Séguin
 Diagnosis

Falcons, parrots, wild birds in zoological or rehabilitation centers

Clinical signs
Epidemiology
Radiology
Endoscopy
Hematology
Biochemistry
Electrophoresis
Cytology, histology
Mycological culture
Serology
PCR ?
 Diagnosis

Falcons, parrots, wild birds in zoological or rehabilitation centers

Clinical signs
Epidemiology
Radiology
Endoscopy
Hematology
Biochemistry
Electrophoresis Juliet Joseph,
Abu Dhabi Falcon Research Hospital
Cytology, histology
Mycological culture
Serology
PCR ?
 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 ?
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
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

Diagnosis

Serology
 detection of galactomannan

• agglutination (Pastorex Aspergillus®)

• polyclonal sandwich ELISA

• monoclonal sandwich ELISA (Platelia Aspergillus®)

 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)

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

Diagnosis

Serology
 detection of galactomannan

Platelia®
Aspergillus

+ -

+ 17 39 56
CIE
- 20 65 85

37 104 141

Le Loch’ et al. ISHAM 2006 (poster))

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 ?)
 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
PCR ? Dahlahausen et al. AAV proc. 2004
 Diagnosis

Turkeys, chickens, quails, ducks, ostriches…

Clinical signs
Epidemiology
Radiology
Endoscopy
Hematology
Biochemistry
Electrophoresis
Cytology, histology
Mycological culture
Serology
Necropsy
 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
 Treatment

Turkeys, chickens, quails, ducks, ostriches…

No treatment !
 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
 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

 References

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