You have a multi-year funded research program designed to measure efficacy of ma

Question

You have a multi-year funded research program designed to measure efficacy of many (hundreds) new anti-microbials that have been developed to treat tuberculosis You will conduct both in vitro and in vivo efficacy tests. Only the candidate anti- microbials which are effective in vitro will be assayed in vivo. Explain your answers fully. 25 points total 8 For the in vivo assays, you will expose guinea pigs via aerosolization (the natural route of exposure for Mycobacterium tuberculosis) to various strains of M. tuberculosis and house them for up to 6 months to assess the efficacy of the anti- microbials. To assess the efficacy, you will necropsy each guinea pig, homogenize the lungs, livers, and spleens of each guinea pig, and spread the homogenates on plates to grow for 30 days. 9 Describe the BSL and PPE you would use when conducting 1) The aerosol exposures 2) Housing and caring for the guinea pigs, 3) The homogenization 4) The plate culture phases of your research.

Explanation / Answer

While handling pathogens in the lab condition at in vitro and in vivo condition there are certain safety measures which has to be followed ,these measures comes under Biosafety level(BSL) and personal protective equipment(PPE).

Biosafety levels deal with the preacautions applied for .regulating laboartory practices without contamination using protective barriers like gloves,lab coat and masks etc.

There are four levels of BSL,I,II,III and IV.

1)Aerosol exposure- Biosafety level III is commonly used for aerosol protection which can be inhaled due to accidental handling and undetected leakage of aerosol to the environment.This can happen during centrifugation,pipetting,blending and sonicating.These harmful particle when inhaled can cause respiratory infections in the person handling the task.

Biosafety III requiremenits are

Double door,self closing entry door

Airation system to direct airflow in and out of the laboratory.

Laboratory away from corridors so that the contaminated particles won't mix with environment.

Autoclave

Post exposure method and vaccination

PPE-Personal [rotective equpment during aerosol exposures are sterilized lab coat,gloves ,face shield and eye glass.

2)Housing and caring for the guinea pig.

Any contacts from infected animals and fluids from their body can also leads to serious hazards.

BSL-Biosafety level III is used for animal handling .

Vaccum and decontamination system

Separate Clothing for enetring to animal shelter.

Anmal shelter separted from main building

PPE-Autoclved clothgs and quipment to handle.

sterilized gloves and eye masks.

Laboratory wastes incinerated.

3)Homogenization of organs

While crushing the isolated tissues and cells from the infected animals may result in transmission of pathogenic microbes.It can happen due to careless handling of the infected animal cells.

BSL II-Biosafety level 2 is associated with handling homogenization of cells and animal organs

Biosafety chamber

Incubator

Refrigerator

Polystyrene treated plastic wears.

PPE-Safety glass to shield eyes from pathogen

Fully covered shoes

autoclaved homogenizer

sterilized clothing,mask and gloves(GMT plus clothing)

4)Handling plate culture

While culturing the bacterial strain in the plates diseases spreding bacteria and viruses can spread to the handler from body fluid or blood.Injestion or inhalation of the particles from careless hadling can leads to lethal hazards.

BS1 is associated with laboratory microbial practices.

GMT(Good microbial techniques are followed)

Laminar air hood

sterilized needles and plates

Minimum traffic

Incinerators

PPE-Well informed handler

Sterile equipments

Sterilized clothing,gloves,eye glass and mask.

No eating,drinking and smoking

using disinfectants for hand wiping.

autoclaved and filtered media

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