Lecture Notes: HEALTH MONITORING in Laboratory Animals

MICROBIOLOGICAL CLASSIFICATION
Lab animals can be classified according to their microbiological status thus:
1. Conventional:- harbour a range of (infectious) microorganisms, kept without
specific preventive hygiene measures and therefore of unknown
microbiological status; used in experiments after undergoing quarantine
2. Specific pathogen free (SPF):- are free from a number of potentially
pathogenic microorganisms. SPF designates which microorganisms are known
not to be present at the time of the last testing
3. Gnotobiotic:- are of defined microflora and are further divided into:
i. Germ-free (GF)- obtained via hysterectomy (re-derivation); are kept
under sterile conditions since they are highly susceptible to infection
ii. Colonisation resistant flora (CRF): GF animals can deliberately be
given flora to provide them with a general resistance to
colonisation/growth of other bacteria (colonisation resistance) and
reduce opportunistic infections. CRF are housed as GF and are often
used to start SPF colonies

The microbiological quality of experimental animals can critically influence

animal welfare and the validity and reproducibility of research data. It is
important that animals are free of agents (latent or apparent) that may interfere
with specific models or projects. Why?
 Some may confound experimental data
 Increase biological and experimental variability
 Cause an increase in animal use
 Some agents are zoonotic
Hence need for a laboratory animal health monitoring program (HM) as an
integral part of quality assurance. Aimed at breeders, users of laboratory animals
and describe essential elements including:
1. choice of agents
2. selection of animals and tissue for testing
3. frequency of sampling
4. common used test methods
5. interpretation of results
6. HM Reporting

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 Requirement of person with sufficient understanding of HM; Important that all
individual who work directly (animal care staff, technicians, researchers) and
indirectly (suppliers) understand the rationale of the HM Program

GENERAL CONSIDERATIONS IN DESIGN OF HM PROGRAM

Animal facilities are structured into microbiological units (MU), defined as a
self-contained microbiological entity, with separate space and traffic for animals,
personnel and materials.
HM Responsible defines the MU and schemes to the use of the unit
 Barrier (Conventional) facility where personnel, equipment and animals
move freely; animals in open cages
 Isolator
i. Microisolation cages- there is direct contacts allowing for horizontal
transmission
ii. Individually Ventilated cage (IVC)- laminar flow cabinet following strict
hygiene measure
The thoroughness of an HM Program must reflect the level of risk that applies to
particular unit and risk it poses to other units
Frequency of monitoring is determined by microbiological characteristics and
prevalence of infectious agents
High risk
i. Frequent introduction of animals eg >1/month
ii. Introduction of animals from different breeding colonies
iii. MU of varying microbiological status close together
iv. Movement of animals within the unit for manipulation and return
v. Access of insects, vermin to animal rooms, feed or bedding
vi. Multipurpose facilities with various kinds of experiments
vii. Frequent entry of research personnel into the MU
viii. Frequent turnover of animal care staff
ix. Shared equipment not easily shared
Low risk
i. Closed breeding colonies
ii. All in – all out system
iii. Occasional introduction of animals
iv. One or few types of experiments

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Choice of agents to monitor
Is determined by several factors:
 Animal health
 Effects on confounding experimental data
 Species specificity- some agents are species specific Sendai virus is a virus
of rodents
 Zoonotic potential eg LCMV
 Prevalence
 Host factors (esp. immune status)
 Desired MU status
 Historical results,

Animals for testing and sampling

 Animals resident within the MU give best results
 Animals in same MU but in several rooms should be sampled from as many
rooms as possible
 Sick and dead animals or their samples should be examined- outcome of
necropsy may prompt an increase in frequency of monitoring
Sentinel animals
 When resident animals are not available or cannot be sampled (study protocol
limitations etc)
 Are exposed to animals of the same species and submitted after adequate
exposure period
 May be either indirect (animals that are exposed to materials soiled by
resident animals e.g. bedding, water, feed) or direct (animals that are placed in
the same cage as resident animals)
 Should be housed in same units for a minimum of 6 weeks before testing using
same husbandry procedures
 Generally should be of the same species as the test animals

Assays and Interpretation

Testing should be under supervision of adequately trained personnel usually
with a degree in microbiology, vet medicine with experience in lab animal science
Accredited laboratory for diagnosis advocated for
Detection of an infectious agent can be by various direct or indirect methods
i. Necropsy- systematic exam of euthanized (or dead) animals for lesions

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 ii. Histopathology should ensue on grossly identified lesion following
published general guidelines
iii. Skin and fur for evidence of parasites
iv. Serology for screening of microorganisms
 Agglutination haemaglutination inhibition tests
 Bead-based assays multiplex bead assays
 ELISA- ubiquitous screening test
 Western blots – good for confirmation due to the high specificity and
sensitivity
v. Molecular methods eg PCR useful for confirmation
vi. Culture techniques for detection of bacterial and fungal agents
Every assay can produce a false-positive and false-negative result hence
prevalence data is important in estimation of predictive value of test results i.e.
how much weight to accord a positive test result

Health Monitoring Report

HM Report is not the a laboratory report
HM Report should be produced by the person incharge of the HM Program and
should adopt a standardized format and must contain the following info
i. MU name and housing condition (barrier, isolator, IVC)
ii. Identification of all species, strains/stocks present within the unit. When
more than one species are reared in same barrier , there should be a HM
Report for each species
iii. Date of the latest test investigation and date of issue of the report
iv. Test frequency for each agent
v. Names of agents for which testing is undertaken to species level
vi. Test method used for each agent and name of testing lab
vii. Results of the latest investigation, and historical data as far as practicable
but at least 18months
viii. Results of pathological examinations recorded as lesions were/were not
observed
ix. A space for comments
x. Name and contact of person responsible for devising the HM Program
xi. Description of the overall HM Progam (link or by cover letter)

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