2010 Mouse Abstracts

Comparative Medicine

Perturbations in Cytokine Gene Expression after Inoculation of

C57BL/6 Mice with Pasteurella pneumotropica

Authors: Patten, Calvin C.1; Myles, Matthew H.2; Franklin, Craig

L.2; Livingston, Robert S.2

Source: Comparative Medicine, Volume 60, Number 1, February 2010 , pp.

18-24(7)

Pasteurella pneumotropica can cause inflammation and abscess formation in

a variety of tissues. Most commonly, P. pneumotropica produces clinical
disease in immunodeficient mice or those concurrently infected with other
pathogens. Because clinical disease is infrequent in immunocompetent mice
harboring P. pneumotropica, some scientists consider it an opportunistic
pathogen with little clinical relevance to biomedical research. However, other
infectious agents, including mouse parvoviruses, mouse rotavirus, and
Helicobacter spp. alter physiologic or biologic responses without causing
clinical signs of illness. We investigated the potential for P. pneumotropica to
modulate the transcription of cytokine genes in immunocompetent mice. In
C57BL/6 mice inoculated oronasally with a minimal colonizing dose of P.
pneumotropica, modest but statistically significant elevations of IL1β, TNFα,
CCL3, CXCL1, and CXCL2 mRNA were detected in mandibular and superficial
cervical lymph nodes at 7 d after inoculation, and upregulation of IL1β mRNA
was detected 28 d after inoculation. These perturbations were not present in
C57/BL6 mice inoculated with heat killed-P. pneumotropica or the related
bacterium Actinobacillus muris. Nasal mucosal cytokine transcription did not
vary significantly in C57BL/6 mice given a high dose of P. pneumotropica.
These data indicate that slight and transient experimental perturbations are
possible in immunocompetent mice colonized with P. pneumotropica.
Knowing the full health status of experimental mice is paramount to avoid
unwanted experimental variables, especially when using exquisitely sensitive
testing methodologies such as those for quantification of gene expression.

The t(14,15) in Mouse Strain CBA/CaH-T(14;15)6Ca/J Causes a Break

in the ADAMTS12 Gene

Authors: Acar-Perk, Bengi1; Bräutigam, Karen2; Grunewald,

Regina1; Schmutzler, Andreas1; Schem, Christian1; Arnold, Norbert
K.1; Jonat, Walter1; Weimer, Jörg1

Source: Comparative Medicine, Volume 60, Number 2, April 2010 , pp. 118-
122(5)
The mouse strain CBA/CaH-T(14;15)6Ca/J carries a homozygous balanced
reciprocal translocation between mouse chromosomes 14 and 15, but the
break points of this translocation have not previously been examined in
detail. Using fluorescent in situ hybridization, we assigned the break point in
14qE3 to a 200-kb region devoid of any known gene. We similarly defined
the break point in 15qA1 to a 27-kb region containing involving ADAMTS12.
The chromosomal break likely is between exons 2 and 3 of ADAMTS12. This
gene encodes a disintegrin and metalloproteinase with thrombospondin
motifs, and this product plays crucial roles in both vascularization and cancer
progression and has been implicated in the development of arthritis. The
CBA/CaH-T(14;15)6Ca/J mouse strain likely is a suitable model for further
examination of the influences of defective ADAMTS12 in various pathologic
processes.

Experimental Infection of Mice with Hamster Parvovirus: Evidence

for Interspecies Transmission of Mouse Parvovirus 3

Authors: Christie, Rachel D.1; Marcus, Emily C.1; Wagner, April

M.1; Besselsen, David G.1

Source: Comparative Medicine, Volume 60, Number 2, April 2010 , pp. 123-
129(7)

Hamster parvovirus (HaPV) was isolated 2 decades ago from hamsters with
clinical signs similar to those induced in hamsters experimentally infected
with other rodent parvoviruses. Genetically, HaPV is most closely related to
mouse parvovirus (MPV), which induces subclinical infection in mice. A novel
MPV strain, MPV3, was detected recently in naturally infected mice, and
genomic sequence analysis indicates that MPV3 is almost identical to HaPV.
The goal of the present studies was to examine the infectivity of HaPV in
mice. Neonatal and weanling mice of several mouse strains were inoculated
with HaPV. Tissues, excretions, and sera were harvested at 1, 2, 4, and 8 wk
after inoculation and evaluated by quantitative PCR and serologic assays
specific for HaPV. Quantitative PCR detected viral DNA quantities that greatly
exceeded the quantity of virus in inocula in multiple tissues of infected mice.
Seroconversion to both nonstructural and structural viral proteins was
detected in most immunocompetent mice 2 or more weeks after inoculation
with HaPV. In neonatal SCID mice, viral transcripts were detected in lymphoid
tissues by RT-PCR and viral DNA was detected in feces by quantitative PCR at
8 wk after inoculation. No clinical signs, gross, or histologic lesions were
observed. These findings are similar to those observed in mice infected with
MPV. These data support the hypothesis that HaPV and MPV3 are likely
variants of the same viral species, for which the mouse is the natural rodent
host with rare interspecies transmission to the hamster.
Effects of Murine Norovirus Infection on a Mouse Model of Diet-
Induced Obesity and Insulin Resistance

Authors: Paik, Jisun1; Fierce, Yvette1; Drivdahl, Rolf1; Treuting, Piper

M.1; Seamons, Audrey1; Brabb, Thea1; Maggio-Price, Lillian2

Source: Comparative Medicine, Volume 60, Number 3, June 2010 , pp. 189-
195(7)

Murine norovirus (MNV) is prevalent in SPF mouse facilities in the United

States, and we currently lack sufficient data to determine whether it should
be eliminated. It is generally accepted that the virus does not cause clinical
symptoms in immuno-competent mice. However, we previously reported that
MNV infection alters the phenotype of a mouse model of bacteria-induced
inflammatory bowel disease in part through its effects on dendritic cells. The
tropism of MNV toward macrophages and dendritic cells makes MNV a
potential intercurrent variable in murine models of macrophage-driven
inflammatory diseases, such as obesity, insulin resistance, and
atherosclerosis. Therefore, we determined whether MNV infection altered
obesity and insulin resistance phenotypes in C57BL/6 mice, a widely used
model of diet-induced obesity. We found that MNV did not alter weight gain,
food intake, and glucose metabolism in this model, but it did induce subtle
changes in lymphoid tissue. Further studies using other models of metabolic
diseases are needed to provide additional information on the potential role
this 'subclinical' virus might have on disease progression in mouse models of
inflammatory diseases.

Male CD81 Knockout Genotype Disrupts Mendelian Distribution of

Offspring

Authors: Mordica, Whitney J.1; Gallagher, Ryan J.1; Kennedy, Jenna

L.1; Chapes, Stephen K.2

Source: Comparative Medicine, Volume 60, Number 3, June 2010 , pp. 196-
199(4)

CD81 is an integral membrane protein in the tetraspanin superfamily that

serves as an adaptor protein. CD81 is also a maternally imprinted gene that
is found in a regulated cluster of genes on mouse chromosome 7. Among
offspring produced from heterozygous breeding pairs, CD81null/null mice grew
at the same rate as CD81+/+ and CD81+/null mice. Because of an inhibition in
sperm-egg fusion, CD81null/null female mice are much less fertile than CD81+/+
and CD81+/null mice. However, no published study has detailed the effect of
the male CD81 genotype on the genotype and sex distribution of offspring.
We set up breeding pairs of heterozygotic (C.129-Cd81tm1 N7) female mice
and male mice with CD81+/null, CD81+/+, or CD81null/null genotypes. The survival
and development of CD81+/null, CD81+/+, and CD81null/null offspring were
monitored and compared. Compared with those of heterozygous male
breeders, CD81null/null pups were born at a less-than-expected ratio from
CD81null/null males. Sex distribution did not differ among pups sired by
CD81null/null compared with CD81+/null mice. The data suggest that the effect of
the CD81null/null paternal genotype on offspring is manifested early in
development or in utero.

Etiopathogenesis of Mandibulofacial and Maxillofacial Abscesses in

Mice

Author: Lawson, Gregory W.1

Source: Comparative Medicine, Volume 60, Number 3, June 2010 , pp. 200-
204(5)

The etiologic agent of mandibulofacial and maxillofacial abscesses in mice is

reportedly coagulase-positive Staphylococcus aureus. Although suggested to
be through the oral cavity, the exact route of entry has not been
documented. Among the clinical cases of mandibulofacial and maxillofacial
abscess we report here, each case that was cultured yielded coagulase-
positive S. aureus. Histologically, all of the abscesses examined were directly
associated with intralesional hair shafts, both vibrissae and pelage, that were
introduced into the submucosa via the maxillary or mandibular molar
gingival sulci. Grossly, a variable amount of hair was imbedded in the lingual,
buccal, or mesial gingival sulci of the maxillary or mandibular molars or both.
Computed tomography revealed that the presence of the hair resulted in
inflammation and resorption of alveolar bone. With these findings, we
propose that mandibulofacial and maxillofacial abscesses are induced by the
mastication and fragmentation of hair ingested during the barbering process.
From the resulting foreign body periodontitis, abscess formation originates at
the maxillary lingual, buccal, or mesial gingival sulci, resulting in infection of
the maxillary molar tooth roots with swelling or rupture through the skin
inferior to the eye, or at the mandibular lingual, buccal, and or mesial
gingival sulci, resulting in infection of the mandibular molar tooth roots and
osteomyelitis with drainage through the skin of the ventral mandible.

Quantitation of Acute Phase Proteins and Protein Electrophoresis in

Monitoring the Acute Inflammatory Process in Experimentally and
Naturally Infected Mice

Authors: Cray, Carolyn1; Besselsen, David G.2; Hart, Jody L.3; Yoon,
David4; Rodriguez, Marilyn5; Zaias, Julia6; Altman, Norman H.5
Source: Comparative Medicine, Volume 60, Number 4, August 2010 , pp.
263-271(9)

Serologic screening for infectious disease in sentinel mice from rodent

colonies is expensive and labor-intensive, often involving multiple assays for
several different infectious agents. Previously, we established normal
reference ranges for the protein fractions of several laboratory strains of
mice by using a commercially available agarose system of protein
electrophoresis. In the current study, we address protein fractionation and
quantitation of acute phase proteins (APP) in mice experimentally infected
with Sendai virus or mouse parvovirus. We further investigate this
methodology by using samples from sentinel mice from colonies with
endemic infection. All study groups showed significant increases in γ
globulins. Various other protein fractions showed mild variable changes;
significant differences were not detected for individual APP. These results
contrast the significant changes observed in APP and protein electrophoresis
by using the standard methods of inducing inflammatory responses through
injection of complete Freund adjuvant or LPS. These present data suggest
that although quantitation of individual APP may not be helpful, γ globulin
levels may reflect infection in laboratory mice and provide a possible adjunct
to traditional screening methods.

Lack of Association of a Spontaneous Mutation of the Chrm2 Gene

with Behavioral and Physiologic Phenotypic Differences in Inbred
Mice

Authors: Ding, Ming1; Arnold, Jennifer1; Turner, Jeremy2; Ramkumar,

Vickram1; Hughes, Larry F.3; Trammell, Rita A.4; Toth, Linda A.5

Source: Comparative Medicine, Volume 60, Number 4, August 2010 , pp.

272-281(10)

The nucleotide substitution C797T in the Chrm2 gene causes substitution of

leucine for proline at position 266 (P266L) of the CHRM2 protein. Because
Chrm2 codes for the type 2 muscarinic receptor, this mutation could
influence physiologic and behavioral phenotypes of mice. Chrm2 mRNA was
not differentially expressed in 2 brain regions with high cholinergic
innervation in a mouse strain that does (BALB/cByJ) or does not (C57BL/6J)
have the mutation. In addition, strains of mice with and without the C797T
point mutation in Chrm2 did not differ significantly in muscarinic binding
properties. Variation across strains was detected in terms of acoustic startle,
prepulse inhibition, and the physiologic effects of the muscarinic agonist
oxotremorine. However, interstrain differences in these measures did not
correlate with the presence of the mutation. Although we were unable to
associate a measurable phenotype with the Chrm2 mutation, assessment of
the mutation on other genetic backgrounds or in the context of other traits
might reveal differential effects. Therefore, despite our negative findings,
evaluation of characteristics that involve muscarinic function should be
undertaken with caution when comparing mice with different alleles of the
Chrm2 gene

Variation in the Gut Microbiota of Laboratory Mice Is Related to Both

Genetic and Environmental Factors

Authors: Hufeldt, Majbritt Ravn1; Nielsen, Dennis S.2; Vogensen, Finn

Kvist2; Midtvedt, Tore3; Hansen, Axel Kornerup4

Source: Comparative Medicine, Volume 60, Number 5, October 2010 , pp.

336-347(12)

During recent years, the composition of the gut microbiota (GM) has received
increasing attention as a factor in the development of experimental
inflammatory disease in animal models. Because increased variation in the
GM might lead to increased variation in disease parameters, determining and
reducing GM variation between laboratory animals may provide more
consistent models. Both genetic and environmental aspects influence the
composition of the GM and may vary between laboratory animal breeding
centers and within an individual breeding center. This study investigated the
variation in cecal microbiota in 8-wk-old NMRI and C57BL/6 mice by using
denaturing gradient gel electrophoresis to profile PCR-derived amplicons
from bacterial 16S rRNA genes. Comparison of the cecal microbiotas
revealed that the similarity index of the inbred C57BL/6Sca strain was 10%
higher than that of the outbred Sca:NMRI stock. Comparing C57BL/6 mice
from 2 vendors revealed significant differences in the microbial profile,
whereas the profiles of C57BL/6Sca mice raised in separate rooms within the
same breeding center were not significantly different. Furthermore, housing
in individually ventilated cages did not lead to intercage variation. These
results show that denaturing gradient gel electrophoresis is a simple tool that
can be used to characterize the gut microbiota of mice. Including such
characterizations in future quality-control programs may increase the
reproducibility of mouse studies.

JAALAS

Effect of Murine Norovirus Infection on Mouse Parvovirus Infection

Authors: Compton, Susan R.1; Paturzo, Frank X.1; Macy, James D.2
Source: Journal of the American Association for Laboratory Animal Science,
Volume 49, Number 1, January 2010 , pp. 11-21(11)

Enzootic infection with mouse parvovirus (MPV) remains a common problem

in laboratory colonies, and diagnosis of MPV infection is complicated by viral
and host factors. The effect of an underlying viral infection on MPV infection
has not previously been investigated. We assessed the effect of murine
norovirus (MNV) infection, the most prevalent infectious agent in laboratory
mice, on MPV shedding, tissue distribution and transmission. Fecal MPV
shedding persisted longer in BALB/c mice infected with MNV 1 wk prior to
MPV infection than in mice infected with MPV only, but transmission of MPV
to soiled-bedding sentinels was not prolonged in coinfected mice. MPV DNA
levels in coinfected BALB/c mice were higher in mesenteric lymph nodes and
spleens at 1 and 2 wk after inoculation and in small intestines at 1 wk after
inoculation compared with levels in mice infected with MPV only. In C57BL/6
mice, fecal shedding was prolonged, but no difference in soiled bedding
transmission or MPV DNA levels in tissues was detected between singly and
coinfected mice. MPV DNA levels in singly and coinfected SW mice were
similar. MPV DNA levels were highest in SW, intermediate in BALB/c and
lowest in C57BL/6 mice. MPV DNA levels in mesenteric lymph nodes of
BALB/c and SW mice exceeded those in small intestines and feces, whereas
the inverse occurred in C57BL/6 mice. In conclusion, MNV infection increased
the duration of MPV shedding and increased MPV DNA levels in tissues of
BALB/c mice.

Anomer-Equilibrated Streptozotocin Solution for the Induction of

Experimental Diabetes in Mice (Mus musculus)

Authors: de la Garza-Rodea, Anabel S.1; Knaän-Shanzer, Shoshan1; den

Hartigh, Jan D.2; Verhaegen, Alphons P.L.2; van Bekkum, Dirk W.1

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 1, January 2010 , pp. 40-44(5)

Streptozotocin is widely used to induce diabetes in laboratory animals

through multiple low-dose or single high-dose intraperitoneal injections.
HPLC analysis has shown that the composition of the solution may change
considerably during the first 2 h after dissolution due to equilibration of the 2
anomers (α and β) of streptozotocin. Because of the drug's alleged instability
in solution, the typical recommendation is to administer streptozotocin within
10 min after dissolution. We compared the induction of diabetes in NOD/SCID
mice by injection of a single high dose of freshly made or anomer-
equilibrated streptozotocin solution. Solutions were prepared from dry
compound containing 85% of the α anomer, which is the more toxic of the 2.
Body weight and nonfasting blood glucose levels were measured weekly for
8 wk. Both solutions induced long-term hyperglycemia, but blood glucose
levels and mortality were higher and damage to pancreatic islands more
pronounced in the mice receiving freshly prepared solution. A small
proportion of mice did not respond in both treatment groups. If stored at 4 °C
in the dark, the anomer-equilibrated solution retains its biologic activity for at
least 40 d; under those conditions the streptozotocin content decreases by
0.1% daily, as determined by HPLC. Anomer-equilibrated streptozotocin
solution has several practical advantages, and we recommend its use as
standard for the induction of experimental diabetes because this practice
may improve reproducibility and comparison of results between different
laboratories.

Suppurative Adenitis of Preputial Glands Associated with

Corynebacterium mastitidis Infection in Mice

Authors: Radaelli, Enrico1; Manarolla, Giovanni2; Pisoni, Giuliano2; Balloi,

Annalisa3; Aresu, Luca4; Sparaciari, Paolo5; Maggi, Adriana6; Caniatti,
Mario2; Scanziani, Eugenio1

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 1, January 2010 , pp. 69-74(6)

Suppuration of the preputial gland in mice occurs as a septic complication of

fight wounds around the external genitalia. Currently reported bacterial
isolates from these lesions are limited to Staphylococcus aureus, Pasteurella
pneumotropica, and Klebsiella oxytoca. In the context of a pilot experiment
aimed at defining the aging phenotype of estrogen receptor β knockout
(BERKO) mice, 2 male mice (1 of the BERKO line and the other from the age-
and sex-matched wild-type control group) were discovered at necropsy to
have preputial gland lesions. In both cases, histopathologic examination
confirmed severe suppuration and abscesses of the preputial glands
associated with systemic reactive (secondary) amyloidosis. Both Gram
staining and Bacillus Calmette-Guérin immunohistochemistry highlighted the
presence of numerous bacillary to rod-shaped bacteria within the preputial
lesions. Subsequent PCR analysis coupled with denaturing gradient gel
electrophoresis identified Corynebacterium mastitidis in the preputial gland
abscesses. This organism is isolated infrequently from the milk of sheep with
subclinical mastitis and was identified as part of the normal microflora of the
human ocular surface. No information regarding the epidemiology and
pathogenesis of C. mastitidis infection in laboratory animals is currently
available, and to our knowledge this report is the first description of C.
mastitidis infection in mice.
Disparities in Ammonia, Temperature, Humidity, and Airborne
Particulate Matter between the Micro-and Macroenvironments of
Mice in Individually Ventilated Caging

Authors: Rosenbaum, Matthew D.1; VandeWoude, Susan2; Volckens,

John3; Johnson, Thomase3

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 2, March 2010 , pp. 177-183(7)

Animal room environmental parameters typically are monitored with the

assumption that the environment within the cage closely mirrors the room
environment. This study evaluated that premise by examining macro- (room)
and microenvironmental (cage) parameters in individually ventilated cages
housing mice with variable amounts of bedding over a period of 17 d without
cage changes. Intracage ammonia levels remained within recommended
human guidelines but were higher than room levels, confirming that
microisolation caging is efficient at preventing ammonia generated from
animal waste from escaping into the room. Humidity and temperature within
cages were consistently higher than room levels. Particles in the room
predominantly consisted of fine particles (diameter less than 2.5 μm),
presumably from the ambient atmosphere; some of these particles were
found in the cage microenvironment. In addition, mouse activity within cages
produced larger particles, and these particles contributed to substantially
higher aerosol mass concentrations within the cage. These findings
demonstrate that, although cage and room environmental parameters differ,
knowledge of room environmental conditions can be used to predict certain
conditions within the cage. This association is relevant in that typical animal
care standard operating procedures rely on room measurements, not
intracage measurements, which arguably are more important for assessing
animal welfare. Further, location and ambient climate can influence particle
concentrations in the room, and consequently within the animal cage,
suggesting local weather patterns and air quality may account for variability
among studies conducted at sites that are geographically divergent.

Clinical Biochemistry Parameters in C57BL/6J Mice after Blood

Collection from the Submandibular Vein and Retroorbital Plexus

Authors: Fernández, Itziar1; Peña, Arantza1; Del Teso, Nahia1; Pérez,

Virginia1; Rodríguez-Cuesta, Juan1

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 2, March 2010 , pp. 202-206(5)
Collection of blood from the submandibular vein allows simple and rapid
processing of many animals without anesthesia and facilitates rapid recovery
with no signs of pain and discomfort in the mice. Here we compared the
submandibular vein and retroorbital plexus blood collection methods, to
determine the potential effect of the sampling technique on several clinical
biochemistry parameters in C57BL/6J mice. We found statistically significant
differences for 8 of the 9 biochemical parameters studied between the 2
blood sampling techniques. Compared with samples collected from the
retroorbital plexus, blood obtained from the submadibular vein had higher
levels of AST, ALT, protein, albumin, triglycerides, total cholesterol, and
creatinine. Glucose values of retroorbital blood were higher than those from
the submandibular vein. Urea levels were similar for both sampling
techniques. Our results demonstrate that the technique used to obtain blood
samples affects parameters commonly used to assess animal health. We
recommend caution when comparing results of biochemical analysis of blood
obtained from the submandibular vein in mice with reference values
obtained by other blood sampling techniques.

Identification of Markers for Imminent Death in Mice used in

Longevity and Aging Research

Authors: Ray, Maria A.1; Johnston, Nancy A.2; Verhulst, Steven3; Trammell,
Rita A.4; Toth, Linda A.5

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 3, May 2010 , pp. 282-288(7)

The goal of this study was to identify objective criteria that would reliably
predict imminent death in aged mice. Male and female ICR mice (age, 8 mo)
were subcutaneously implanted with an identification chip for remote
measurement of body temperature. Mice then were weighed and monitored
regularly until spontaneous death occurred or until euthanasia was
administered for humane reasons. Clinical signs that signaled
implementation of euthanasia included inability to walk, lack of response to
manipulation, large or ulcerated tumors, seizures, and palpable hypothermia.
In mice that died spontaneously, gradual weight loss was the most frequent
and earliest sign of imminent death. Hypothermia developed during the 2 wk
prior to death. Slow or labored breathing were observed in about half of the
mice before death. A composite score of temperature × weight can be used
to provide an objective benchmark to signal increased observation or
euthanasia of individual mice. Such assessment may allow the collection of
terminal tissue samples without markedly altering longevity data, although
application of this criterion may not be appropriate for all studies of
longevity. Timely euthanasia of mice based on validated markers of imminent
death can allow implementation of endpoints that alleviate terminal distress
in aged mice, may not significantly affect longevity data, and can permit
timely collection of biologic samples.

Eradication of Helicobacter spp. by Using Medicated Diet in Mice

Deficient in Functional Natural Killer Cells and Complement Factor D

Authors: del Carmen Martino-Cardona, Maria1; Beck, Sarah E.1; Brayton,

Cory1; Watson, Julie1

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 3, May 2010 , pp. 294-299(6)

A commercial 4-drug diet has shown promise in eradicating Helicobacter spp.

from rodents; however, its effectiveness in immunocompromised mice is
unknown. This study evaluated the efficacy of this treatment in eradicating
Helicobacter spp. from mice deficient in functional natural killer cells (Cd1−/−)
or complement factor D (Df−/−). Cd1−/− mice naturally infected with H.
hepaticus with or without H. rodentium were fed either control or medicated
diet for 8 wk followed by 4 wk on control diet. Fecal samples were PCR-
evaluated for Helicobacter spp. before mice began treatment and then every
2 wk thereafter for 12 wk. The same experimental design was repeated for
eighteen 9- to 21-wk-old Df-/- mice naturally infected with H. bilis with or
without H. rodentium. All Df-/- mice and 8- to 21-wk-old Cd1−/− mice ceased
shedding Helicobacter spp. after 2 wk of treatment and remained negative
throughout the study. In contrast, the Cd1−/− mice that were 24 wk or older
shed Helicobacter spp. for the first 8 wk but tested negative at 10 and 12 wk.
All treated animals had enlarged ceca and gained less weight than control
untreated mice, and 6 of 7 treated Cd1−/− male mice developed mild portal
fibrosis. These findings show that within 2 wk of treatment, the 4-drug diet
eradicated H. hepaticus and H. rodentium from young Cd1−/− mice and H.
bilis and H. rodentium from Df−/− mice, but eradication of established
infections in Cd1−/− mice required 8 wk of treatment.

Vacuum-Cleaner Noise and Acute Stress Responses in Female

C57BL/6 Mice (Mus musculus)

Authors: Jensen, Kelly1; Hahn, Nina E.1; Palme, Rupert2; Saxton,

Katherine3; Francis, Darlene D.4

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 3, May 2010 , pp. 300-306(7)

Audiogenic stress is a well-documented phenomenon in laboratory rodents.

Despite the recommendation in the Guide for the Care and Use of Laboratory
Animals to consider noise a concern in the animal facility, only a small body
of literature empirically addresses the effects of facility noise on laboratory
rodents, particularly mice. The objective of this study was to determine
whether facility noise generated by a vacuum cleaner induces an acute
stress response in a commonly used strain of laboratory mouse under
common housing conditions. In each of 2 experiments, 10 young adult,
female C57BL/6Cr mice were exposed for 1 h to noise produced by a vacuum
cleaner, and 10 control mice were not. In the first experiment, fecal samples
were collected to measure concentrations of fecal corticosterone metabolites
just before and 2, 4, 6, 8, 10, 14, 24, and 32 h after noise exposure. In the
second experiment, stress-sensitive behavioral tests were performed 2 d
before, immediately after, and 24 h after noise exposure. Physiologic and
behavioral measurements indicated that vacuum cleaner noise did not cause
an acute stress response in the noise-exposed mice but may have affected
the diurnal variation of their corticosterone levels. These findings could
contribute to the development of best practices in noise-control protocols for
animal facilities.

Evaluation of a Commercial Colorimetric Fecal Dipstick Assay for the

Detection of Helicobacter hepaticus Infections in Laboratory Mice

Authors: Freebersyser, Julie E.1; Drake, Michael T.1; Riley, Lela K.1; Myles,
Matthew H.1; Livingston, Robert S.1

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 3, May 2010 , pp. 312-315(4)

Mice used in biomedical research typically are tested for the presence of
Helicobacter spp., including Helicobacter hepaticus. Here we evaluated the
ability of a commercially available colorimetric Helicobacter dipstick assay to
detect H. hepaticus in experimentally and naturally infected mice, with use
of a Helicobacter PCR assay as the 'gold standard' test. None of the fecal
samples from experimentally infected A/JCr mice (n = 12) tested positive for
Helicobacter by the colorimetric dipstick test. In naturally infected A/JCr and
C57BL/6 mice, 11% (1 of 9) and 30% (3 of 10) of fecal samples, respectively,
tested positive for Helicobacter by the colorimetric dipstick assay. In these 3
groups of H. hepaticus-infected mice, statistically fewer mice tested positive
by the colorimetric dipstick test than by PCR. The colorimetric Helicobacter
dipstick assay had an overall diagnostic sensitivity of 13%, diagnostic
specificity of 94%, and analytical sensitivity of 108 H. hepaticus cfu/mL. As
currently formulated, the colorimetric dipstick assay had high specificity but
lacked sensitivity for detecting H. hepaticus infections in 2 strains of mice
commonly used in research, thereby limiting its utility as a diagnostic
screening test for H. hepaticus infections in research mice.

Euthanasia by CO2 Inhalation Affects Potassium Levels in Mice

Authors: Traslavina, Ryan P.1; King, Edward J.2; Loar, Andrew S.2; Riedel,
Elyn R.3; Garvey, Michael S.4; Ricart-Arbona, Rodolfo3; Wolf, Felix R.3; Couto,
Suzana S.3

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 3, May 2010 , pp. 316-322(7)

We and others frequently have noted serum potassium levels of 8.0 ± 0.85
mEq/L or greater in laboratory mice; this concentration has even been
published as the upper limit of a 'normal' reference range. However, if bone
fide, this potassium concentration would be incompatible with life in all
species. We investigated conditions frequently encountered in the research
setting to distinguish artifactual from true hyperkalemia. Variables evaluated
included site of collection, time allowed for clot formation before serum
separation, time elapsed between collection and analysis of samples
collected in a serum separator tube, precollection method of anesthesia, and
euthanasia technique. Serum potassium was measured from 75 C57BL/6NTac
10-wk-old female mice and divided into at least 5 mice per variable. Animals
were euthanized by exsanguination immediately after terminal CO2 or
ketamine-xylazine (KX) administration. Mice euthanized with CO2 had higher
mean serum potassium (7.0 ± 0.5 mEq/L) and range serum potassium (6.0
to 8.1 mEq/L) than did KX-treated mice. CO2 inhalation resulted in
significantly lower blood pH (6.9 ± 0.1), higher pCO2 (153.3 ± 38.8 mm Hg),
and higher lactate levels (3.9 ± 0.9 mmol/L) than did KX anesthesia followed
by exsanguination. These results suggest that antemortem respiratory
acidosis from CO2 administration causes artifactual hyperkalemia in mice.
Therefore, blood collection under KX anesthesia is preferable over CO2
inhalation to obtain accurate potassium values from mice.

Evaluation of Hydration and Nutritional Gels as Supportive Care

after Total-Body Irradiation in Mice (Mus musculus)

Authors: Moccia, Krinon D.1; Olsen, Cara H.2; Mitchell, Jennifer

M.1; Landauer, Michael R.1

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 3, May 2010 , pp. 323-328(6)

Concern regarding the potential for radiation exposure from accidents or

nuclear and radiologic terrorism is increasing. The purpose of this study was
to determine whether the addition of minimal supportive care consisting of
hydration or nutritional gels could be used to reduce mortality in mice
exposed to 60Co γ-radiation. Male CD2F1 mice received 0, 8.50, or 9.25 Gy
60
Co at a dose rate of 0.6 Gy/min. These groups were further divided into 3
treatment groups that—in addition to pelleted food and water—received no
supportive care, hydration gel, or nutritional gel. Overall survival, mean
survival time, consumption of pelleted food and gel, and body weight were
recorded for 30 d. Radiation caused dose-dependent decreases in overall
survival, consumption of pelleted food and supplemental gel, and body
weight. However, at each radiation dose (0, 8.50, 9.25 Gy), the type of
supportive care did not modify overall survival, mean survival time, or
changes in body weight. These results demonstrate that hydration and
nutritional gels were not effective methods of supportive care after high-dose
total body irradiation in mice.

A Spoonful of Sugar Helps the Medicine Go Down: A Novel Technique

to Improve Oral Gavage in Mice

Authors: Hoggatt, Amber F.1; Hoggatt, Jonathan2; Honerlaw,

Meghan3; Pelus, Louis M.2

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 3, May 2010 , pp. 329-334(6)

Oral gavage is a common route of precise oral dosing for studies in rodents.
Complications including tracheal administration, esophageal trauma, and
aspiration are common and usually related to animal resistance to the
procedure, and the stress induced by oral gavage can be a confounding
variable in many studies. The taste of sucrose conveys a pacifying and
analgesic effect in newborns, whereas sour solutions can induce the swallow
reflex in humans that are dysphagic. We hypothesized that precoating a
gavage needle with sucrose or citrate (or both) would pacify mice and induce
them to swallow, reducing the stress and complications associated with the
technique. To validate this hypothesis, we quantitated time to passage,
stress-related behavioral reactions to the procedure, and plasma
corticosterone levels in mice after precoating gavage needles with water,
sucrose, citrate, sucrose and citrate, or sodium chloride prior to oral gavage.
Precoating needles with sucrose reduced the time to passage, decreased
observable stress-related reactions to the procedure, and maintained plasma
corticosterone levels similar to those in ungavaged control mice. Coating
needles with water, sucrose and citrate, or citrate had no beneficial effects
on these parameters. Our findings describe a novel, validated technique that
measurably decreases signs of stress and thereby improves animal welfare
during oral gavage. Furthermore, the use of sucrose may be a valuable tool
to refine other minor or nonsurgical procedures in the field of laboratory
animal research.

Short-Term Storage and Transport at Cold Temperatures of 2-Cell

Mouse Embryos Produced by Cryopreserved Sperm
Authors: Takeo, Toru1; Kondo, Tomoko1; Haruguchi, Yukie1; Fukumoto,
Kiyoko1; Nakagawa, Yoshiko1; Takeshita, Yumi1; Nakamuta,
Yuko1; Tsuchiyama, Shuuji1; Shimizu, Norihiko2; Hasegawa, Takanori3; Goto,
Motohito4; Miyachi, Hitoshi5; Anzai, Masayuki6; Fujikawa, Rie7; Nomaru,
Koji8; Kaneko, Takehito1; Itagaki, Yoshiaki9; Nakagata, Naomi10

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 4, July 2010 , pp. 415-419(5)

At refrigerated temperatures, mouse embryos can maintain developmental

ability for short periods. Previously, we succeeded in transporting vitrified
and warmed 2-cell mouse embryos while maintaining developmental ability
at refrigerated temperatures for 50 h. Transport of nonfrozen embryos is an
easier and more useful means of exchanging genetically engineered mice
between laboratories than is transport of cryopreserved embryos. Here we
examined the developmental ability of transported 2-cell embryos that were
produced through in vitro fertilization using cryopreserved sperm. Results
show that 2-cell embryos produced by cryopreserved sperm can develop into
blastocysts after cold storage for 24, 48, and 72 h. Transported 2-cell
embryos produced by cryopreserved sperm yielded a favorable number of
pups in all of the receiving laboratories after transport lasting 48 to 52 h. In
summary, cold storage and transport of 2-cell embryos derived from
cryopreserved sperm at refrigerated temperatures provides a novel means of
transporting genetically engineered mice as an alternative to the transport of
cryopreserved embryos and sperm.

Refinements in the Cryopreservation of Mouse Ovaries

Authors: Sztein, Jorge1; Vasudevan, Kuzhalini2; Raber, James3

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 4, July 2010 , pp. 420-422(3)

Here we describe a new technique for cryopreserving mouse ovaries by

using 0.5-mL straws. One advantage of this method is that it uses the same
controlled-rate freezer and programming routinely used for the
cryopreservation of mouse embryos. Using a 0.5-mL French straw loaded in
the same way as for embryo freezing (for example, the one-step dilution
method) with 1 M sucrose as an osmotic buffer and 2 M propylene glycol as
the cryoprotectant containing the ovary sample, we further standardized the
2 methodologies. Applying this technique, 11 ovarian halves were
cryopreserved in straws and stored under liquid nitrogen. Straws containing
the frozen ovarian halves were thawed in a water bath at room temperature
and the recovered ovaries orthotopically implanted into 11 recipient female
mice; 8 of the 11 frozen ovarian halves resulted in functional ovaries. The
73% pregnancy rate resulted in a total of 53 pups born, of which 38 (72%)
were generated from cryopreserved ovaries. Ovarian cryopreservation has
been demonstrated to be a valid option for banking mouse genetic
resources. Unlike frozen embryos, cryopreservation of ovarian tissue
preserves haploid gametes. Despite this limitation, ovarian cryopreservation
is the only technique that can be used to preserve oocytes from aged or
problematic breeders. This advantage is especially important in situations
where the only males available in the line are infertile, aged, or problematic
breeders.

The Effects of Perioperative Analgesia on Litter Size in Crl:CD1(ICR)

Mice Undergoing Embryo Transfer

Authors: Goulding, David R.1; Myers, Page H.2; Goulding, Eugenia

H.3; Blankenship, Terry L.2; Grant, Mary F.2; Forsythe, Diane B.2

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 4, July 2010 , pp. 423-426(4)

The objective of this study was to evaluate the effect on litter size of 2
analgesics used perioperatively during mouse embryo transfer surgery. Day
2.5 pseudopregnant CD1 mice (n = 96) were divided equally into 2 analgesic
treatment groups and a saline control group. Each mouse received a single,
subcutaneous dose of buprenorphine hydrochloride (0.1 mg/kg), flunixin
meglumine (2.5 mg/kg), or saline immediately after induction of anesthesia
with 2.5% isoflurane. Each mouse then was prepared for aseptic surgery.
Blastocysts had previously been collected from C57BL/6NCrl female mice
that were synchronized and superovulated by using pregnant mare serum
gonadotropin and human chorionic gonadotropin and mated with
C57BL/6NTac male mice 3.5 d before collection. Viable blastocysts were
pooled, and 8 were selected arbitrarily and transplanted into the right
uterine horn of each pseudopregnant CD1 mouse. Mice were monitored
throughout pregnancy, and the number of pups at birth was documented. No
statistically significant difference was found between the 3 groups. These
results indicate that perioperative analgesic treatment with buprenorphine or
flunixin in the CD1 mouse undergoing embryo transfer is not associated with
increased embryonic loss.

Serologic Prevalence of MPV1 in Mouse Strains in a Commercial

Laboratory Mouse Colony Determined by Using VP1 Antigen

Authors: Filipovska-Naumovska, Emilija1; Abubakar, Salleh M.2; Thompson,

Martin J.1; Hopwood, Deborah2; Pass, David A.2; Wilcox, Graham E.3

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 4, July 2010 , pp. 437-442(6)
A mouse parvovirus (designated MPV1f) was identified in a commercial
laboratory mouse colony in Australia. The infection had not been detected by
using an rNS1 parvovirus ELISA antigen even though the virus was
genetically similar to other MPV1 variants reported previously. A recombinant
biotinylated protein based on a truncated VP1 protein of the MPV1 strain was
produced and used as antigen for ELISA and Western immunoblots to detect
virus infection and determine the seroprevalence of infection in a colony of
approximately 45,000 mice. Antibody-positive mice were detected in 8 of 11
rooms sampled, indicating that infection was widespread in the facility.
Antibody was detected in 16.2% of 1161 sera obtained from 20 strains of
mice. Seroprevalence varied among mouse strains, suggesting genetic
variation in the susceptibility of mice to MPV1 or in their antibody response
to infection, as has been reported previously in experimentally infected mice.
Seroprevalence was high in some inbred strains, including DBA/2JArc and the
random-bred strains Hsd:NIH and Arc:Arc(s). Antibody was not detected
inC57BL/6J strains, and BALB/c strains showed low seroprevalence of MPV1f.

Strain- and Age-Associated Variation in Viral Persistence and

Antibody Response to Mouse Parvovirus 1 in Experimentally
Infected Mice

Authors: Filipovska-Naumovska, Emilija1; Thompson, Martin J.1; Hopwood,

Deborah2; Pass, David A.2; Wilcox, Graham E.3

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 4, July 2010 , pp. 443-447(5)

The effect of mouse strain and age at infection on viral replication and
concurrent antibody response to mouse parvovirus 1 (isolate MPV1f) was
evaluated for 305 d after inoculation in 4 strains of mice. The results
confirmed previous reports that mouse strain and age at infection are
significant factors in viral persistence and antibody development and
detection. Randombred Arc:Arc(s) mice originally bred from CD1 stock
inoculated as juveniles (4 wk) or adults (8 wk) developed persistent viral
infection for 152 d after inoculation and an antibody response that persisted
for 295 d. Mice of C57BL/6J background inoculated as juveniles had
detectable viral DNA in large intestinal content and tissues for 24 d after
inoculation and an antibody response that persisted for 288 d. However, viral
DNA was not detected in tissues of C57BL/6J mice inoculated as adults,
although an antibody was detected for 111 d after inoculation; these results
suggest probable viral replication in adult C57BL/6J mice but at levels below
the limits of detection. BALB/cArc mice inoculated as juveniles or adults had
detectable virus DNA in tissues for 108 to 242 d after inoculation, but no
antibody was detected. Similarly, BALB/c-Foxn1nu/Arc mice had detectable
levels of viral DNA in tissues for 98 to 131 d but no measurable antibody. The
difficulty of detecting antibody in mice with a BALB/c background indicates
they are unsuitable for routine surveillance of MPV1f infection.

Spontaneous Staphylococcus xylosus Infection in Mice Deficient in

NADPH Oxidase and Comparison with Other Laboratory Mouse
Strains

Authors: Gozalo, Alfonso S.1; Hoffmann, Victoria J.2; Brinster, Lauren

R.2; Elkins, William R.3; Ding, Li4; Holland, Steven M.4

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 4, July 2010 , pp. 480-486(7)

Staphylococcus xylosus typically is described as a nonpathogenic common

inhabitant of rodent skin. Reports of S. xylosus as a primary pathogen in
human and veterinary medicine are scarce. Here we report 37 cases,
affecting 12 strains of laboratory mice, of spontaneous infections in which S.
xylosus was isolated and considered to be the primary pathogen contributing
to the death or need for euthanasia of the animal. Infection with S. xylosus
was the major cause of death or euthanasia in 3 strains of mice deficient in
the production of phagocyte superoxide due to defects in NADPH oxidase.
NADPH-oxidase-deficient mice (n = 21) were most susceptible to
spontaneous S. xylosus infections. The infections were characterized by
abscesses and granulomas in soft tissues, with bacterial migration to internal
organs (primarily regional lymph nodes and lungs and, to a lesser degree,
muscle, bone, and meninges). In contrast, 9 strains of phagocyte-superoxide-
producing mice (n = 16) also had S. xylosus infections, but these were
largely confined to eyelids, ocular conjunctiva, and skin and rarely involved
other tissues or organs. Because exhaustive bacterial culture and isolation
may not be performed routinely from mouse abscesses, S. xylosus infections
may be underdiagnosed. S. xylosus should be considered in the differential
diagnosis in laboratory mice with abscesses and other skin lesions. This
report expands the range of mouse strains and tissues and organs
susceptible to spontaneous S. xylosus infection and compares the pathology
among various mice strains.

Treatment and Eradication of Murine Fur Mites: I. Toxicologic

Evaluation of Ivermectin-Compounded Feed

Authors: Ricart Arbona, Rodolfo J.1; Lipman, Neil S.1; Riedel, Elyn R.2; Wolf,
Felix R.1

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 5, September 2010 , pp. 564-570(7)
Fur mite outbreaks remain a persistent problem in laboratory mouse
colonies. All currently published treatment methods are labor-intensive,
expensive, or unreliable. During a recent outbreak with Myobia musculi and
Myocoptes musculinus in a large colony (approximately 30,000 cages), we
developed a feed-based treatment regime in which ivermectin was the active
ingredient. Rodent feed was compounded with 3 different concentrations of
ivermectin (12, 24, and 48 ppm) and γ-irradiated. Postcompounding analysis
revealed loss of ivermectin during manufacturing, but the remaining drug
was stable for at least 6 mo. In an 8-wk toxicity study in a C57BL/6NTac
mouse breeding colony, ad-libitum feeding of the 3 diets yielded estimated
doses of 1.3, 2.7, and 5.4 mg/kg. Adult mice lacked adverse clinical effects,
except that 1 of the 144 mice in the 48-ppm group developed tremors and
ataxia and was euthanized. No significant differences between doses were
revealed by CBC, serum chemistry, body weight, or gross necropsy. Plasma
drug concentrations plateaued at a dose-dependent level 7 to 10 d after
initiation of treatment and decreased to undetectable levels 6 to 9 d after its
discontinuation. Fertility of the P0 generation was unaffected. Pup mortality
was higher in the 24- and 48-ppm groups, reaching 100% at the higher dose.
Animals exposed to ivermectin as neonates had normal weaning weights, but
mice receiving 24-ppm feed had lower adult weights. Our results indicate
that using feed containing 12 ppm ivermectin (estimated ingested dose, 1.3
mg/kg) was safe in a C57BL/6NTac breeding colony.

Treatment and Eradication of Murine Fur Mites: II. Diagnostic

Considerations

Authors: Ricart Arbona, Rodolfo J.1; Lipman, Neil S.1; Wolf, Felix R.1

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 5, September 2010 , pp. 583-587(5)

Fur mites are a persistent problem in contemporary laboratory mouse

colonies. We conducted several studies to evaluate fur mite diagnostic
methodologies and interpretation of results. Retrospective analysis of test
results from sentinel mice exposed to soiled bedding collected from colonies
infested with Myobia musculi and Myocoptes musculinus revealed the skin
scrape test to be more reliable than pelt examination, provided that both the
head and dorsal thoracolumbar regions were sampled. To assess their
diagnostic accuracy, 3 commercial laboratories were sent positive control
slides containing mites, mite parts, or eggs in sets of slides containing
diagnostic skin scrapings in varying ratios. Laboratory B correctly identified
the positive control slide. Laboratory A identified 1 of 3 positive control
slides, whereas laboratory C failed to identify both positive control slides
submitted. To determine the time required for a mouse to shed its entire hair
coat, fur of Crl:CD1(ICR), BALB/cAnNCrl, and Crl:CFW(SW) albino mice was
dyed black and the presence of dyed fur evaluated monthly for 8 mo. Limited
dyed hair was still present at 8 mo; therefore, finding eggs or egg casings
many months after treatment cessation does not necessarily imply treatment
failure. To evaluate the effectiveness of soiled bedding sentinels for detection
of fur mites in a mite-infested colony, we exposed naïve mice to varying
amounts (100%, 50%, 25%, 2.5%, and 0%) of soiled bedding in clean
bedding. As little as 2.5% soiled bedding resulted in detection of a positive
sentinel within a 2-mo period.

Resident Bacterial Flora in the Skin of C57BL/6 Mice Housed under

SPF Conditions

Authors: Tavakkol, Zarry1; Samuelson, Derrick2; deLancey Pulcini,

Elinor2; Underwood, Robert A.1; Usui, Marcia L.1; Costerton, J
William3; James, Garth A.2; Olerud, John E.1; Fleckman, Philip1

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 5, September 2010 , pp. 588-591(4)

Research in cutaneous biology frequently involves models that use mice

housed in SPF conditions. Little information is available concerning the
species of bacteria that normally inhabit the skin of these mice. The aim of
this study was to characterize the bacterial skin flora of mice housed under
SPF conditions. Skin biopsies from C57BL/6 mice under normal and surgically
prepped conditions were both cultured and analyzed by using DNA extraction
and sequencing. The species isolated most commonly from culture were
staphylococci. Coagulase-negative staphylococci were isolated more
frequently than was Staphylococcus aureus. Molecular sequencing yielded
several additional organisms not found by culture. Overall, culturing of
isolates yielded 14 species of bacteria, and molecular sequencing identified
another 6 species. Investigators conducting cutaneous research in mouse
models should aware of the cutaneous bacterial flora present on these mice.

Noise in a Laboratory Animal Facility from the Human and Mouse

Perspectives

Authors: Reynolds, Randall P.1; Kinard, Will L.2; Degraff, Jesse J.1; Leverage,
Ned3; Norton, John N.1

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 5, September 2010 , pp. 592-597(6)

The current study was performed to understand the level of sound produced
by ventilated racks, animal transfer stations, and construction equipment
that mice in ventilated cages hear relative to what humans would hear in the
same environment. Although the ventilated rack and animal transfer station
both produced sound pressure levels above the ambient level within the
human hearing range, the sound pressure levels within the mouse hearing
range did not increase above ambient noise from either noise source. When
various types of construction equipment were used 3 ft from the ventilated
rack, the sound pressure level within the mouse hearing range was increased
but to a lesser degree for each implement than were the sound pressure
levels within the human hearing range. At more distant locations within the
animal facility, sound pressure levels from the large jackhammer within the
mouse hearing range decreased much more rapidly than did those in the
human hearing range, indicating that less of the sound is perceived by mice
than by humans. The relatively high proportion of low-frequency sound
produced by the shot blaster, used without the metal shot that it normally
uses to clean concrete, increased the sound pressure level above the
ambient level for humans but did not increase sound pressure levels above
ambient noise for mice at locations greater than 3 ft from inside of the cage,
where sound was measured. This study demonstrates that sound clearly
audible to humans in the animal facility may be perceived to a lesser degree
or not at all by mice, because of the frequency content of the sound.

Assessment of Carprofen and Buprenorphine on Recovery of Mice

after Surgical Removal of the Mammary Fat Pad

Authors: Adamson, Trinka W.1; Kendall, Lon V.1; Goss, Sherri1; Grayson,
Kevin2; Touma, Chadi3; Palme, Rupert4; Chen, Jane Q.5; Borowsky, Alexander
D.5

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 5, September 2010 , pp. 610-616(7)

The purpose of this study was to determine the level of pain elicited by
mammary fat pad removal surgery and the effects of postoperative
analgesics on recovery. Female FVB mice were anesthetized, and mammary
fat pad removal was performed. After surgery, mice received carprofen,
buprenorphine, a combination of carprofen and buprenorphine, or saline
treatment. Additional mice received anesthesia but no surgery or treatment.
Food and water intake, body weight, wheel running activity, and a visual
assessment score were recorded daily for 4 d after surgery and compared
with presurgical findings. Corticosterone metabolites in fecal samples were
analyzed at 12 and 24 h postsurgically and compared with baseline values.
All surgical groups had significantly decreased food intake at 24 h, with a
return to baseline by 48 h. The combination treatment resulted in a
significantly decreased water intake and body weight at 24 h. All surgical
groups had significantly decreased wheel running activity at 24 h only. The
visual assessment scores indicated mild pain for all surgical groups, with the
buprenorphine treated mice showing the highest pain index scores, as
compared with nonsurgical controls. Fecal corticosterone metabolite levels
did not differ significantly between any of the groups or across time. The
parameters used in this study did not indicate that administration of these
analgesic regimens improved recovery as compared with that of saline-
treated mice. Care should be taken when using visual assessment scores to
evaluate pain in mice, given that analgesics may have side effects that
inadvertently elevate the score.

Treatment and Eradication of Murine Fur Mites: III. Treatment of a

Large Mouse Colony with Ivermectin-Compounded Feed

Authors: Ricart Arbona, Rodolfo J.1; Lipman, Neil S.1; Wolf, Felix R.1

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 5, September 2010 , pp. 633-637(5)

We determined the efficacy of ivermectin-compounded feed against fur

mites in mice and describe its use to eradicate mites in vivaria holding
approximately 30,000 cages. C57BL/6NCrl mice infested with Myobia musculi
and Myocoptes musculinus were treated with ivermectin-compounded feed
(approximate ingested dose, 1.3 mg/kg) for 1, 4, or 8 consecutive weeks.
Regardless of treatment duration, all treated mice, as well as contact
sentinels, remained free of fur mites for as long as 21 wk after treatment. No
adverse effects were observed. Subsequently, facility-wide treatment was
implemented in an attempt to eradicate fur mites from 3 vivaria housing
approximately 120,000 mice. Medicated feed was provided for 8 wk to
ensure that all cages and mice were treated. A single investigative group
reported adverse effects in their colony 4 wk after treatment was initiated;
mortality was attributed to ivermectin toxicity after an intracranial injection
at 1 d of age. Naïve pups were unaffected. No other adverse effects were
noted. Approximately 14,500 skin scrape samples were evaluated during the
12-mo posttreatment surveillance period. All samples were negative for
mites. To our knowledge, this is the first report of successful eradication of
fur mites from a mouse colony of this large size.

Effect of Fenbendazole on Three Behavioral Tests in Male C57BL/6N

Mice

Authors: Gadad, Bharathi S.1; Daher, João P.L.2; Hutchinson, Eric

K.3; Brayton, Cory F.3; Dawson, Ted M.4; Pletnikov, Mikhail V.5; Watson, Julie6

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 6, November 2010 , pp. 821-825(5)
Pinworms are highly contagious parasites of laboratory rodents that often are
treated with fenbendazole. To our knowledge, the effect of fenbendazole at
therapeutic dosages on behavioral tests in mice has not been evaluated.
Here we studied 6-wk-old male C57BL/6N mice. We compared the behavior
of control mice (fed regular diet) with 3 groups of mice treated with dietary
fenbendazole. Treatment groups were 4 wk of fenbendazole, 2 wk of
fenbendazole followed by 2 wk of regular diet, and 2 wk of regular diet
followed by 2 wk of fenbendazole. At the end of dietary treatment all groups
were tested by open field for central, peripheral and vertical activity;
elevated plus maze for anxiety; and rotarod for motor ability and then
evaluated by clinical pathology and selected histopathology. Treated and
control groups showed no differences in open field or elevated plus maze
testing, histopathology, or clinical pathology. However mice treated for 4 wk
with fenbendazole or 2 wk of fenbendazole followed by 2 wk regular diet
stayed on the rotarod for shorter periods than did controls, and mice treated
with 2 wk of regular diet followed by 2 wk fenbendazole showed a trend
toward shorter rotarod times. In light of this study, we suggest that open
field and elevated plus maze testing is unlikely to be affected by 4 wk
fenbendazole treatment in male C57BL/6 mice; however, behavioral tests of
motor ability such as rotarod tests may be affected during and for at least 2
wk after fenbendazole treatment.

Efficacy of Soaking in 70% Isopropyl Alcohol on Aerobic Bacterial

Decontamination of Surgical Instruments and Gloves for Serial
Mouse Laparotomies

Authors: Keen, Jessica N.1; Austin, Marykay2; Huang, Li-Shan3; Messing,

Susan3; Wyatt, Jeffreyd2

Source: Journal of the American Association for Laboratory Animal Science,

Volume 49, Number 6, November 2010 , pp. 832-837(6)

Rodent surgeries in biomedical research facilities are often performed in

series. This practice presents many challenges to maintaining aseptic
technique between animals. Here, we examined using soaking in 70%
isopropyl alcohol for aerobic bacterial decontamination of surgical
instruments and gloves used in a series of as many as 10 mouse laparotomy
surgeries. These surgeries were performed on mice that were euthanized
immediately prior to the procedure. Instruments and gloves were cultured
before and after each procedure to determine the presence of aerobic
bacterial contamination. To assess the efficacy of the decontamination
protocol, culture results were grouped by procedure and then paired (before
soak and after soak) for analysis using McNemar test at an α level of 0.05. In
addition, by using the Fisher exact test, this modified aseptic method was
compared with strict aseptic technique, for which autoclaved instruments
and sterile surgical gloves were used for each procedure. In this study, we
observed that the modified aseptic technique using 70% isopropyl alcohol
soaks pre- vented aerobic bacterial contamination of instruments and gloves
for as many as 5 mice.