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Jacques Guillot, Guillaume Le Loch, Pascal Arné, Françoise Féménia, René Chermette Avianaspergillosis UMR INRA, AFSSA, ENVA, UPVM 956, Biologie Moléculaire.

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Presentation on theme: "Jacques Guillot, Guillaume Le Loch, Pascal Arné, Françoise Féménia, René Chermette Avianaspergillosis UMR INRA, AFSSA, ENVA, UPVM 956, Biologie Moléculaire."— Presentation transcript:

1 Jacques Guillot, Guillaume Le Loch, Pascal Arné, Françoise Féménia, René Chermette Avianaspergillosis UMR INRA, AFSSA, ENVA, UPVM 956, Biologie Moléculaire et Immunologie Parasitaires et Fongiques, Ecole Nationale Vétérinaire dAlfort, 94704 Maisons-Alfort Cedex, France

2 Introduction Aspergillosis is considered as one of the most important infectious diseases in birds early 1800s: first observations early 1800s: first observations scaup duck, jay, bullfinch, bustard and several swans 1898: first cases in turkey poults (Lignières & Petit in France) 1898: first cases in turkey poults (Lignières & Petit in France) Since then, cases in a very wide range of avian species… Since then, cases in a very wide range of avian species…

3 5 key questions… Why are birds more susceptible than mammals ? In which circumstances does aspergillosis occur in birds ? Is it possible to make an early diagnosis in birds ? Is it possible to treat or prevent avian aspergillosis ? What do the main clinical signs and lesions look like ?

4 Susceptibility of birds Birds are much more susceptible to aspergillosis than mammalian species Environmental contamination by Aspergillus spp. Environmental contamination by Aspergillus spp.

5 Susceptibility of birds Environmental contamination by Aspergillus spp. Environmental contamination by Aspergillus spp. Avian anatomy and physiology Avian anatomy and physiology 40-43°C40-43°C Birds are much more susceptible to aspergillosis than mammalian species lungs / air sacs

6 Susceptibility of birds Environmental contamination by Aspergillus spp. Environmental contamination by Aspergillus spp. Avian anatomy and physiology Avian anatomy and physiology Avian immunology Avian immunology Lack of resident macrophages in airways and air sacs Heterophils without peroxydation Birds are much more susceptible to aspergillosis than mammalian species

7 Epidemiology Aspergillus fumigatus / Aspergillus flavus, Aspergillus niger… Overgrowth Specific virulence factors ? humidity, dampness drying period …

8 Epidemiology a 600 m 2 confinement building in the Center of France a 600 m 2 confinement building in the Center of France a flock comprising 4500 turkeys a flock comprising 4500 turkeys females slaughtered at the age of 12 w and males at the age of 16 w 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 An epidemiological survey was conducted in France

9 Epidemiology Lair-Fulleringer et al. Poultry Science 2006

10 Epidemiology 5 males 5 females 10 healthy chicks, 23 isolates, 1 unique genotype Lair-Fulleringer et al. J. Clin. Microbiol. 2003

11 Epidemiology 9 healthy turkeys, 55 isolates, 17 genotypes Lair-Fulleringer et al. J. Clin. Microbiol. 2003

12 Epidemiology 2 carcass condemnations, 36 isolates, 2 genotypes Lair-Fulleringer et al. J. Clin. Microbiol. 2003

13 Epidemiology 362 air samples, 134 isolates 53 genotypes 16 week-sampling period Lair-Fulleringer et al. Poultry Science 2006

14 Epidemiology Aspergillus fumigatus / Aspergillus flavus, Aspergillus niger… Overgrowth Specific virulence factors ? Species Avian strains Avian strains Individual susceptibility ? Individual susceptibility ? Age Sex Stress Other micro-organisms Other micro-organisms Toxicosis Therapeutics captive environment handlingmigration… mycotoxins ? corticosteroids turkeys, quails, raptors, penguins, parrots, waterfowl

15 Clinical signs and lesions Different avian species Different epidemiological situations Different avian species Different epidemiological situations Many clinical signs Acute aspergillosis = inappetance, depression, polydipsia, polyuria, dyspnoea, cyanosis = sometimes, sudden death without any signs Acute aspergillosis = inappetance, depression, polydipsia, polyuria, dyspnoea, cyanosis = sometimes, sudden death without any signs Chronic aspergillosis = signs are dependent on the area of invasion change in voice respiratory stridor exercice intolerance ataxia, torticollis, seizures conjunctivitis, keratitis beak malformation… Chronic aspergillosis = signs are dependent on the area of invasion change in voice respiratory stridor exercice intolerance ataxia, torticollis, seizures conjunctivitis, keratitis beak malformation… fatal evolution

16 Clinical signs and lesions ENVA Kaminski et al. air sacculitis Aspergillosis of eggs ENVA

17 Clinical signs and lesions Chute pneumonia meningo- encephalitis Séguin arthritis Séguin uveitis Séguin

18 Diagnosis Falcons, parrots, wild birds in zoological or rehabilitation centers Clinical signs Epidemiology RadiologyEndoscopy Hematology Serology Biochemistry Electrophoresis Mycological culture PCR ? Cytology, histology

19 Diagnosis Clinical signs Epidemiology Radiology Endoscopy Hematology Serology Juliet Joseph, Abu Dhabi Falcon Research Hospital Falcons, parrots, wild birds in zoological or rehabilitation centers Biochemistry Electrophoresis Mycological culture PCR ? Cytology, histology

20 Hematology Diagnosis Clinical signs Epidemiology Radiology Endoscopy Serology Falcons, parrots, wild birds in zoological or rehabilitation centers Biochemistry Electrophoresis Mycological culture PCR ? Cytology, histology a promising approach ?

21 Diagnosis Serology Immunologically, birds respond to Aspergillus infection in the same way as mammals and a type I response appears most beneficial 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 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 Serological tests that may be used in birds include the detection of specific antibodies or fungal antigens

22 Diagnosis Serology detection of specific antibodies 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 High prevalence of seropositivity in captive penguins Low prevalence of seropositivity in wild birds

23 Diagnosis Serology detection of galactomannan agglutination (Pastorex Aspergillus®) monoclonal sandwich ELISA (Platelia Aspergillus®) polyclonal sandwich ELISA

24 Diagnosis Serology detection of galactomannan Cray et al. ISHAM 2006 (poster P-0011) 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 !

25 Diagnosis Serology detection of galactomannan Arca-Ruiba et al. Vet. Rec. 2006 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

26 Diagnosis Serology detection of galactomannan Le Loch et al. ISHAM 2006 (poster)) Platelia® Aspergillus +- CIE +173956 -206585 37104141

27 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 ?)

28 Serology Hematology Diagnosis Clinical signs Epidemiology Radiology Endoscopy Falcons, parrots, wild birds in zoological or rehabilitation centers Biochemistry Electrophoresis Mycological culture PCR ? Cytology, histology Hardy et al. AAV proc. 2003 Dahlahausen et al. AAV proc. 2004

29 Diagnosis Clinical signs Epidemiology Turkeys, chickens, quails, ducks, ostriches… HematologyRadiologyEndoscopy Serology Biochemistry Electrophoresis Mycological culture Necropsy Cytology, histology

30 Treatment Falcons, parrots, wild birds in zoological or rehabilitation centers Amphothericin B FlucytosineKetoconazoleItraconazoleTerbinafine Voriconazole (Langhofer, AAV proc. 2004) Radiography, endoscopy or serology for the follow up + Supportive therapy + Surgical debridement of the lesions

31 Treatment No treatment ! Turkeys, chickens, quails, ducks, ostriches…

32 Prevention Reduction of fungal contamination Chimioprevention ? Vaccination with a heat-killed culture filtrate preparation has been reported to reduce mortality in ducks and waterfowl Reduction of stress Vaccination Richard (1984) reduced mortalities by 50% in turkey poults vaccinated with germinated A. fumigatus conidia

33 Conclusions « Avian aspergillosis » = not a single entity but a complex of several diseases… = not a single entity but a complex of several diseases… importance of prophylactic procedures importance of prophylactic procedures poor prognosis poor prognosis need for specific avian models of aspergillosis need for specific avian models of aspergillosis variable epidemiological situations variable epidemiological situations several diagnostic tools required several diagnostic tools required

34 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 Loch, 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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