A Nebulized Intra-tracheal Rat Model of Invasive Pulmonary Aspergillosis
Animal models are particularly useful for the study of many infectious diseases, including those caused by fungi. Invasive pulmonary aspergillosis is most frequently studied in mouse models. We present here an animal model of this disease based on underno
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1. Introduction Aspergillosis is an avian disease with major economic repercussions. Diverse models of this infection were initially developed in bird species: chickens, ducks, turkeys, guinea fowl, and quails. However, towards the end of the twentieth century, the increasing importance of invasive forms of this disease in human medicine (1) has led to increasing experimentation in mammals. Rabbits, guinea pigs, cows, and monkeys have all been studied, but the majority of research is based on rodent models, particular those involving mice (2–4). However, the small size of the mouse makes some protocols difficult, particularly those requiring the repeated sampling of biological material. In this context, rat models present a number of advantages. Rats are inexpensive, easy to manipulate, are large enough to allow for repeated blood sampling and also allow for in vivo imaging. The model presented here is based on previous rat model work (5, 6) but is a new one never used before or elsewhere to our
Alexandra C. Brand and Donna M. MacCallum (eds.), Host-Fungus Interactions: Methods and Protocols, Methods in Molecular Biology, vol. 845, DOI 10.1007/978-1-61779-539-8_36, © Springer Science+Business Media, LLC 2012
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knowledge. The animals are fed a low protein diet and are immunocompromised by cyclophosphamide treatment. Once experimental neutropenia has been established, Aspergillus fumigatus spores are administered by intra-tracheal nebulizer with a Microsprayer IA-1B®. Infection leads to the reproducible development of an experimental pulmonary disease within 3–6 days, with a global mortality of 80% after 10 days. The results of Aspergillus antigen determinations on blood samples correlate with anatomical and pathological data, consistent with published findings (7). Similarities between the clinical and biological aspects of the rat model and those encountered in the human disease confirm the value of this invasive pulmonary aspergillosis model (8).
2. Materials 2.1. Animal Rearing and Handling
1. Rats: male Sprague-Dawley rats (Harlan; see Note 1), aged 6–8 weeks, weight 200–225 g (see Note 2). 2. Protected environment housing; ventilated racks with controlled humidity and temperature (Iffa Credo) containing type 3 cages (50 cm × 25 cm × 20 cm) (Tecniplast; see Note 3). 3. Standard diet (Special Diets Services): 18% Crude protein, 6.6% ash, 3.7% crude fibre, crude 3.1% lipid, 1% calcium, 0.7% phosphorus, 0.39% methionine, 0.26% sodium, 0.18% magnesium, 20,000 IU/kg vitamin A, 2,000 IU/kg vitamin D3, 74 IU/kg vitamin E, 15 mg/kg copper. Standard diet should be stored in a granular form in 12.5 kg bags, in cool, dry conditions. 4. Precision balance (max = 1,000 g; d = 0.01 g). 5. Low protein diet (Safe): 20% maize starch, 9.2% casein, 55.2% sucrose, 5.4% maize oil, 5.5% cellulose, 1% vitamin mix, 1.3% mineral mix, 2.1% calcium phosphate, 0.1% dl-methionine, 0.2% calcium carbonate. The low protein diet should be stored as powder and/or granules, in vacuum packaging, in cool
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