Intranasal Mycobacterial Infection in Mice

Animal studies were carried out in strict accordance with the Amsterdam protocol on animal protection and welfare, the Directive 2010/63/EU of the European Parliament and the Council of 22 September 2010 on the protection of animals used for scientific purposes and the French Decree 2013-118. The protocol was approved by the Minister of Higher Education and Research after favorable opinion of the Ethics Committee (CEEA Nord-Pas de Calais/INSERMU1019 n° 00579.01 from 23/07/2014). All efforts were made to minimize suffering of the animals. Six-weeks-old female BALB/c mice (Janvier) were challenged with mycobacterial strains via the intranasal route with 10 μl/nostril. Challenge suspensions were adjusted in order to obtain an inhaled dose of approximately 1,000 CFU/lungs. At day 0, 28, and 49 post-infection, lungs from euthanized mice were collected and homogenized by using an MM300 apparatus (Qiagen) and 2.5-mm diameter glass beads. Ten-fold serial dilutions of lung homogenates were plated on 7H11/OADC agar medium and CFUs enumerated, as previously described (Palucci et al., 2016 (link)).

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Camassa S., Palucci I., Iantomasi R., Cubeddu T., Minerva M., De Maio F., Jouny S., Petruccioli E., Goletti D., Ria F., Sali M., Sanguinetti M., Manganelli R., Rocca S., Brodin P, & Delogu G. (2017). Impact of pe_pgrs33 Gene Polymorphisms on Mycobacterium tuberculosis Infection and Pathogenesis. Frontiers in Cellular and Infection Microbiology, 7, 137.

Publication 2017

BALB/c mice MM300 apparatus

Agar Animals Balb c mice Dilutions European Female Infection Lung Mice Minister Mycobacterial Research ethics committee Strains

Corresponding Organization : Università Cattolica del Sacro Cuore

Other organizations : Center for Infection and Immunity of Lille, Institut Pasteur de Lille, Université de Lille, Centre Hospitalier Universitaire de Lille, University of Sassari, Istituto Nazionale per le Malattie Infettive Lazzaro Spallanzani, University of Padua

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Variable analysis

independent variables

Mycobacterial strains

dependent variables

Colony-forming units (CFUs) enumerated in lung homogenates

control variables

Animal age (6-weeks-old)
Animal species (female BALB/c mice)
Challenge route (intranasal)
Challenge dose (approximately 1,000 CFU/lungs)
Time points of lung collection (day 0, 28, and 49 post-infection)

controls

No positive or negative controls were explicitly mentioned in the protocol.

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