` Marc Khalaf

Assignment (Week 4) – Marc Khalaf

1. A Report is NOT required for the (3) Readings in the Week 4 folder.
However, please answer the following questions:

a. What does the acronym OLAW stand for?

Office of Laboratory Animal Welfare

b. According to the “Guide,” what is the recommended temperature in F° for

mice as listed in Table 3.1 (pg. 48)?
68-79

c. According to the “Guide,” what is one possible complication of

environmental enrichment for mice (pg. 57-58)?
Some scientists have raised concerns that environmental enrichment may compromise
experimental standardization by introducing environmental variability, adding not only diversity
to the animals’ behavioral repertoire but also variation to the animals’ responses to
experimental treatments (e.g., Bayne 2005; Eskola et al. 1999; Gärtner 1999; Tsai et al. 2003).

It can also lead to aggression and social stress.

d. According to the “Guide,” what is the recommended minimum floor area in

in2 for 5 mice that weigh 25 grams (seep pg. 62). Note: some
institutions may have more stringent regulations
12*5=60

e. According to the “Guide,” what is one possible complication for nude or

hairless mice if one provides cotton nestlets as an environmental
enrichment (see pg. 73).
Cotton nestlets may lead to conjunctivitis

f. According to the paper by Barthold, what did Strong do to embellish the

gene pool of many of his mouse lines, including the C3H strain?
Because of the limited number of DBA mice, Strong embel- lished the gene pool for many of
his mouse lines with wild mice captured from a nearby pigeon coop at Cold Spring Harbor
(thus the agouti coloration of C3H mice, which were derived from wild mice crossed with
albinos and dilute brown nonagoutis).

g. What does the acronym EDIM stand for?

Epizootic diarrhea of infant mice

h. What is the causative agent of EDIM?

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Experimental mice developed posterior paresis Mouse Rotavirus.

i. Who documented the first bacterial disease (“pseudotuberculosis”) in

mice?
D. Kutscher (1894)

j. What does IFA stand for? How is it performed?

Immunofluorescence assay. Detects cross- reactive nonstructural antigens.

Immunofluorescent Assay. If a mouse is infected with a viral or

bacterial pathogen, it will generate antibodies specific to the pathogen.
Its serum can be tested for the presence of this pathogen-specific
antibody using IFA  cells or tissues infected with the pathogen is
fixed to a microscope slide. The test mouse serum is added,
incubated, then washed off. If pathogen-specific antibodies are
present they will bind to the pathogen and remain on the slide. Then a
2° antibody generated against mouse antibodies AND labeled with a
fluorescent dye is added, incubated, then washed off. This antibody
will bind to any mouse pathogen-specific antibody on the slide. The
slide is then examined under a fluorescent microscope. If mouse
pathogen-specific antibodies are present, they will glow under the UV
light.

k. Approximately what percentage of the mouse genome is composed of

retroelements (e.g., retroviruses, retrotransposons)?
37% of the total laboratory mouse genome.

l. What is the Schaedler “cocktail” and what was it used for?

Schaedler created a “cocktail” of selected enteric bacteria from normal mice, and commercial
breeders used these organisms to “associate” their rodent colonies with defined microflora

m. Give 3 common mouse mutations that have been linked to spontaneous

retrovirus integrations.
Rodless retina (rd1, the recessive allele associated with reti- nal degeneration) in Strong’s C3H
mice, hairless mutation (hr), and the dilute (d) coloration of Little’ s
(Lathrop’ s, Tyzzer’s, and Strong’s) DBA mice.

n. Why should you be cautious if your colleague states that his/her mice are
raised under “specific pathogen free (SPF)” conditions?
“Specific pathogen free” is a notoriously nebulous phrase, and meaningless without specifying
the pathogens. Furthermore, the credibility of test results is vital, but these are affected by a
variety of factors such as the testing labo- ratory, test method, sample size, animal age, and
sentinel or indigenous animals.
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2. Go to http://jaxmice.jax.org/health/index.html and select “Animal

Health & Genetic Quality” in menu on the left. Go to “Health Status Reports.”
a. What is the difference between a Standard vs. Maximum Barrier?
Standard barrier: static or individually ventilated caging; Pasteurized or sterilized feed;
change into sterilized clothing and shoes; gloves, mask, cap; restricted entry
Maximum barrier: sterilized individually ventilated caging; sterilized feed and drinking water;
air or wet shower; change into sterilized clean room scrubs, smock and shoes; gloves, air hat
or mask, cap and face shield

b. Examine the Health Reports for G1210 / G1230. What excluded pathogen
was identified in 2009 (previous 12 months)?
Helicobacter spp.

c. Compare the health reports of AX6 (Standard Barrier) vs. AX11 (Maximum
Barrier). What is the difference in the response to identifying
Klebsiella spp.?
The health reports of AX6 responded in significantly decreasing the number of cases (from 40
to around 8 per period) while the health reports of AX 11 did not change significantly over the
time period.

In the Standard Barrier, no attempt has to be made to eliminate

Klebsiella. In the Maximum Barrier, an attempt is made to eliminate
Klebsiella by testing and culling infected mice.

d. Health reports are always retrospective, never prospective. If you ordered

mice from AX8 in Oct 2010 do you have to worry that your mice may
be infected with Klebsiella spp?
No you don’t because all the mice from Oct 2010 will have similar results in the
examination for Klebsiela spp infection.

YES! There is a history of Klebsiella in this barrier and any test is always of
a subset of mice. The barrier may turn positive in the next test result. I
would test any mice I receive for Klebsiella if it is a pathogen you do not want
in your colony.

e. Go to http://jaxmice.jax.org/health/index.html and select “Index of

JAX® Mice & Room Numbers.” Select “View list sorted by room
number.” How many lines of mice are housed in AX9.- 8 strains

AX9A/J000646 AX9BALB/cJ000651 AX9C3H/HeOuJ000635 AX9C57BL/10J000665

AX9CBA/J000656 AX9DBA/1LacJ001140 AX9DBA/2J000671 AX9MRL/MpJ000486
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f. Stanford does not allow direct importation of mice from JAX Research
Animal Facilities. Go to http://jaxmice.jax.org/health and select
room MGL-373. Which pathogens that are normally excluded in the
Maximum barriers are allowed in this Low barrier?
Helicobacter spp, Mouse norovirus (MNV), Pasteurella pneumotropica, Opportunistic protozoa
(e.g., Giardia, Spironucleus)

3. Go to http://info.criver.com/health/

a. Select “Health Monitoring of Laboratory Rodents.” How often do they

screen for Mouse Parvovirus? Mouse Hepatitis Virus? Lymphocytic
Choriomeningitis Virus? Parasites?
Every 4 weeks. Every 4 weeks. Every 13 weeks. Every 13 weeks.

b. Select “Index by Strain.” What pathogens were identified in the outbred

stock CD-1 mouse from Hollister in area H43? What action is taken
by Charles River Labs upon finding these opportunistic pathogens?
Which barrier would your order your CD-1 mice from, H43 or H48?
Staph. Aureus, and Beta Strep. Grp B.
H48 because no pathogens.

4. Examine the Health Reports of Harlan’s production facility at Livermore, CA

(go to http://www.harlan.com/research_models_and_services
and select Health and Genetic Monitoring Reports then select Livermore
(237). What bacterium was identified in mice in 237 within the last 18
months?
None for all mutant, outbred and inbred mice.

Proteus mirabilis, Staphylococcus aureus