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Mycobacterium tuberculosis in a red-crowned parakeet (Cyanoramphus novaezelandiae).

Abstract: An adult female red-crowned parakeet (Cyanoramphus novaezelandieae) was presented for necropsy and histopathologic evaluation. The bird had died after exhibiting lameness, weight loss, respiratory signs, and hemoptysis. Postmortem radiographs revealed lesions in the diaphysis of the left femur and soft-tissue opacities in the lungs. At necropsy, the muscles of the left femur were pale and swollen, white-yellow small nodules were visible in lungs and air sacs, and the liver and other coelomic organs appeared pale. On histologic examination, areas of necrosis in the lungs were extensively infiltrated with acid-fast positive bacilli surrounded by macrophages, epithelioid cells, and giant cells. Acid-fast bacilli were also present in the left leg muscle and in granulomas in the liver, kidneys, and intestine. Fungal hyphae associated with a Splendore-Hoeppli phenomenon were visible in the left leg muscle. Mycobacterium tubereulosis was confirmed by polymerase chain reaction testing performed on muscle samples. Results were indicative of infection with M tuberculosis complicated by mycotic myositis. The disease in avian species is of zoonotic importance, and infected birds may be a useful sentinel for human infection.

Key words: mycobacteriosis, Mycobacterium tuberculosis, polymerase chain reaction, avian, red-crowned parakeet, Cyanoramphus novaezelandiae

Clinical Report

A 3-year-old female red-crowned parakeet (Cyanoramphus novaezelandiae), yellow mutation, was presented for necropsy and histopathologic evaluation. The parrot had originated from a facility in the Netherlands and had a history of chronic lameness and a swollen left leg. The bird had been treated with antibiotics (enrofloxacin) and nonsteroidal anti-inflammatory drugs (meloxicam) for 5 days with no improvement. Its clinical condition had progressively deteriorated, with episodic diarrhea, respiratory rattles, and hemoptysis, which became more evident before death. The carcass was chilled and sent to the Unit of Veterinary Pathology of the University of Messina for necropsy.

Postmortem whole-body radiographs were taken before necropsy. On the ventrodorsal view, an aggressive lesion, characterized by osteolysis, expansion of the cortex with smooth margins, swelling of the surrounding soft tissue, was visible in the left femur. Multiple small radiopaque areas were also observed in the lungs.

At necropsy, the bird was in poor body condition. Several single or confluent white-yellow nodules were visible in the lungs. The walls of the air sacs appeared thickened and opaque with deposits of white-yellow debris. Serosal membranes were thickened. The liver was decreased in size and pale, and all other coelomic organs appeared pale. The left leg was enlarged, and the muscles of the femoral area appeared swollen (Fig 1) and were pale pink on the cut surface. The femoral bone appeared thin. Samples of lungs, kidneys, liver, heart, intestines, and the left leg were collected, fixed in 10% buffered formalin, and routinely processed. Tissue sections were embedded in paraffin wax, cut into 5-[micro]m-thick sections, and stained with hematoxylin and eosin, periodic acid-Schiff, Ziehl-Neelsen, Congo red, Lillie, and Grocott stains. DNA was extracted from paraffin-embedded thigh muscle and lung tissues by xylene treatment and by use of QIAamp Blood and Tissue Kit (QIAGEN s.p.a., 20151, Milano, Italy). The target DNA for amplification was a 580-bp fragment of the IS6110, an insertion sequence-like element currently used to identify members of the Mycobacterium tuberculosis complex. (1) As a positive control, Mycobacterium bovis (B/29292) DNA was included. The resulting polymerase chain reaction (PCR) products were compared by WU BLAST software (Advanced Biocomputing, St Louis, MO, USA) with the genomic data showed on GenBank (National Center for Biotechnology Information, at the BX842581 accession number.


On histologic examination, necrotic foci, surrounded by macrophages, giant cells, and epithelioid cells, were visible in the lungs. On Ziehl-Neelsen-stained sections, numerous positive acid-fast bacilli were visible in the cytoplasm of epithelioid and giant cells. Granulomas with acid-fast bacilli were also seen in the liver, kidneys, intestines, and muscle tissue. Trabecular osteomyelitis was observed in histologic sections of the left leg, and acid-fast positive bacilli were visible in the thigh muscle (Fig 2). Grocott staining revealed numerous septate hyphae in these same areas of muscle; some fungal hyphae were surrounded by intensely eosinophilic material in asteroid- or star-like configurations characteristic of the Splendore-Hoeppli phenomenon.

Polymerase chain reaction testing revealed an amplified product of expected size. It was purified from gel and sequenced by Big Dye terminator kit version 3.1 (Applied Biosystems, Life Technologies Corporation, Carlsbad, CA, USA) and analyzed by 3130 genetic analyzer (Applied Biosystems). We used universal bacterial primers for ribotyping, and the specificity of the amplified was investigated by sequencing. The obtained sequence was compared by WU BLAST software, and the bacteria were identified as M tuberculosis. No other primers were used to identify other more typical Mycobacterium species because the sequence obtained indicated a 100% homology with IS6110 element of M tuberculosis.


In this report, we describe mycobacteriosis caused by M tuberculosis infection in a domestic red-crowned parakeet. Avian mycobacteriosis is most commonly caused by Mycobacterium avium, Mycobacterium intracellulare, and Mycobacterium genavense, whereas infections caused by M tuberculosis, Mycobacterium fortuitum, and M bovis are uncommon or rare. (2-4) Infection with M tuberculosis has been reported in many animals, including primates, cattle, sheep, dogs, cats, and elephants? Infections caused by acid-fast bacteria have been widely reported in domestic poultry, (6,7) as well as in zoo birds. (8) Mycobacterial infections have not been reported in red-crowned parakeets. (9) Localized skin lesions associated with mycobacteriosis are uncommon in birds and have been associated with M tuberculosis. (10)


Ubiquitous species of fungi, such as Aspergillus species, are commonly associated with pathologic lesions in birds. Although these fungi can be a primary cause of respiratory infection, they are often found as opportunistic pathogens in conjunction with other disease processes or in immunosuppressed animals. (11) In humans, aspergillosis is considered an immunologically mediated lung disease and is commonly found in immunocompromised patients, such as those with acquired immunodeficiency syndrome, cancer, or organ transplants. (12,13) Although we could not identify the fungal species by PCR, muscle lesions caused by Aspergillus species have been reported recently in humans (14) as well as in birds. (10) Invasive fungal lesions associated with different muscle pathologies, such as neoplasia and granulomatous inflammation, need accurate methods of differential diagnosis. (15,16)

The Splendore-Hoeppli phenomenon is the formation of intensely eosinophilic material around microorganisms such as fungi, bacteria, and parasites or inert substances. Although the exact nature of this reaction is unknown, it is thought to be a localized immunologic response to an antigen-antibody precipitate. This phenomenon is not typical in avian pathology and represents an unusual finding, here described only around fungi.

In the case we describe, M tuberculosis was considered the primary pathogen because of the diffuse presence of the organism in lesions and the associated pathologic changes. Our results also confirmed the validity of the PCR for the postmortem diagnosis and identification of M tuberculosis on paraffin-embedded tissues.

The disease in avian species is of zoonotic importance, and infected birds may be a useful sentinel for human infection. This has been suggested in other reports of M tuberculosis in an African grey parrot (Psittacus erithacus), (17) experimentally infected budgerigars (Melopsittacus undulatus), (18) a blue-fronted Amazon parrot (Amazona aestiva aestiva), (2,19) a green-winged macaw (Ara chloroptera), (4,20) and a canary (Serinus canaria), (2) as well as in unspecified psittacine birds. (21) Although mycobacteriosis is frequently reported in birds, infection caused by M tuberculosis is not typical in avian disease. Unfortunately, we could not identify the source of infection in this bird by the anamnestic data.


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Giovanni Lanteri, DVM, Fabio Marino, DVM, PhD, Stefano Reale, BSc, Fabrizio Vitale, DVM, Francesco Marci, DVM, and Giuseppe Mazzullo, DVM

From Dipartimento di Sanita Pubblica Veterinaria, Facolta di Medicina Veterinaria, Polo Universitario dell'Annunziata, 98168 Messina, Italy (Lanteri, Marino, Macri, Mazzullo); and Istituto Zooprofilattico Sperimentale della Sicilia "A. Mirri," Via Gino Marinuzzi 3, 90129 Palermo, Italy (Reale, Vitale).
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Article Details
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Title Annotation:Clinical Reports
Author:Lanteri, Giovanni; Marino, Fabio; Reale, Stefano; Vitale, Fabrizio; Macri, Francesco; Mazzullo, Gius
Publication:Journal of Avian Medicine and Surgery
Article Type:Report
Geographic Code:4EUIT
Date:Mar 1, 2011
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