Long Term Chronic <em>Pseudomonas aeruginosa</em> Airway Infection in Mice

Marcella Facchini; Ida De Fino; Camilla Riva; Alessandra Bragonzi

doi:10.3791/51019

Crônica de Longo Prazo Pseudomonas aeruginosa Infecção das vias aéreas em ratos

JoVE Journal
Immunology and Infection

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Immunology and Infection

Long Term Chronic Pseudomonas aeruginosa Airway Infection in Mice

Crônica de Longo Prazo Pseudomonas aeruginosa Infecção das vias aéreas em ratos

Full Text

24,040 Views

15:43 min

March 17, 2014

DOI: 10.3791/51019-v

Marcella Facchini^1,2, Ida De Fino^1,2, Camilla Riva^1,2, Alessandra Bragonzi¹

¹Infections and Cystic Fibrosis Unit, Division of Immunology, Transplantation and Infectious Diseases,San Raffaele Scientific Institute, ²Italian Cystic Fibrosis Research Foundation

Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This article describes the agar-beads method to establish persistent long-term chronic Pseudomonas aeruginosa airway infection in a mouse model. The method allows for stable bacterial load persistence, facilitating research on chronic infections.

Key Study Components

Area of Science

Microbiology
Infectious Diseases
Animal Models

Background

Pseudomonas aeruginosa is a common pathogen in chronic lung infections.
Establishing a reliable animal model is crucial for studying these infections.
Current methods may not provide long-term infection stability.
The agar-beads method aims to overcome these limitations.

Purpose of Study

To develop a method for inducing chronic Pseudomonas aeruginosa infections in mice.
To evaluate the persistence of bacterial load over time.
To assess the effectiveness of this method compared to existing techniques.

Methods Used

Culturing Pseudomonas aeruginosa bacterial strain.
Embedding bacteria in agar beads using agar and mineral oil.
Injecting agar bead suspension into the trachea of mice.
Collecting bronchoalveolar lavage fluid and lung tissue for analysis.

Main Results

Successful establishment of chronic infections in a high percentage of mice.
Stable bacterial load maintained for up to one month.
Effective detection of bacterial content in lung tissues.
Comparison with existing methods shows improved stability.

Conclusions

The agar-beads method is a reliable approach for chronic infection studies.
This method enhances the understanding of Pseudomonas aeruginosa infections.
Future research can build on this model for therapeutic testing.

Frequently Asked Questions

What is the agar-beads method?

It is a technique to establish chronic Pseudomonas aeruginosa infections in mice using agar-embedded bacteria.

Why is this method important?

It allows for stable long-term infections, which are crucial for studying chronic lung infections.

How long can the bacterial load be maintained?

The bacterial load can be maintained for up to one month.

What are the advantages of this method?

It provides a high percentage of successful chronic infections compared to existing methods.

What can be analyzed from the infected mice?

Bronchoalveolar lavage fluid and lung tissues can be analyzed for bacterial content.

Is this method applicable to other pathogens?

While this study focuses on Pseudomonas aeruginosa, the method may be adapted for other pathogens.

Descreve-se o método de ágar-esferas de estabelecer infecção das vias aéreas persistente a longo prazo crônica Pseudomonas aeruginosa no modelo de mouse.

O objetivo geral deste procedimento é estabelecer infecção crônica persistente de longo prazo das vias aéreas de aerógenos de pseudomonas em camundongos. Isso é feito primeiro cultivando a cepa bacteriana de aérgeno pseudomonas selecionada. O segundo passo é incorporar bactérias em pequenas esferas de ágar misturando bactérias com ágar e óleo mineral a 50 graus Celsius e, em seguida, resfriando a mistura a quatro graus Celsius sob agitação lenta e contínua.

Em seguida, o óleo mineral é removido por várias lavagens com PBS e uma alíquota das esferas de ágar é homogeneizada e banhada em placas de ágar Para determinar o conteúdo bacteriano, a etapa final é injetar alguns microlitros da suspensão de esferas de ágar na traqueia de camundongos para estabelecer uma infecção pulmonar crônica por aérgeno de pseudomonas. Em última análise, o fluido de lavagem broncoalveolar e os pulmões podem ser recuperados em diferentes momentos do desafio e plaqueados em placas de ágar para detectar a porcentagem de camundongos cronicamente infectados e a carga bacteriana nas ondas de rádio dos camundongos sobreviventes. A principal vantagem desses métodos aga bits sobre os métodos existentes é que eles permitem uma infecção crônica estável a longo prazo em uma alta porcentagem de camundongos, levando à persistência de uma carga bacteriana estável por até um mês.

View the full transcript and gain access to thousands of scientific videos