2010 mouse journal abstracts

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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, CraigL.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 ina variety of tissues. Most commonly, P. pneumotropica produces clinicaldisease in immunodeficient mice or those concurrently infected with other

pathogens. Because clinical disease is infrequent in immunocompetent miceharboring P. pneumotropica, some scientists consider it an opportunisticpathogen with little clinical relevance to biomedical research. However, otherinfectious agents, including mouse parvoviruses, mouse rotavirus, andHelicobacterspp. alter physiologic or biologic responses without causingclinical signs of illness. We investigated the potential for P. pneumotropica tomodulate the transcription of cytokine genes in immunocompetent mice. InC57BL/6 mice inoculated oronasally with a minimal colonizing dose ofP.pneumotropica, modest but statistically significant elevations ofIL1, TNF,CCL3, CXCL1, and CXCL2 mRNA were detected in mandibular and superficialcervical lymph nodes at 7 d after inoculation, and upregulation ofIL1 mRNA

was detected 28 d after inoculation. These perturbations were not present inC57/BL6 mice inoculated with heat killed-P. pneumotropica or the relatedbacteriumActinobacillus muris. Nasal mucosal cytokine transcription did notvary significantly in C57BL/6 mice given a high dose ofP. pneumotropica.These data indicate that slight and transient experimental perturbations arepossible in immunocompetent mice colonized with P. pneumotropica.Knowing the full health status of experimental mice is paramount to avoidunwanted experimental variables, especially when using exquisitely sensitivetesting 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 theADAMTS12 Gene

Authors: Acar-Perk, Bengi1; Brutigam, Karen2; Grunewald,Regina1; Schmutzler, Andreas1; Schem, Christian1; Arnold, NorbertK.1; Jonat, Walter1; Weimer, Jrg1

Source:Comparative Medicine, Volume 60, Number 2, April 2010 , pp. 118-122(5)
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The mouse strain CBA/CaH-T(14;15)6Ca/J carries a homozygous balancedreciprocal translocation between mouse chromosomes 14 and 15, but thebreak points of this translocation have not previously been examined indetail. Using fluorescent in situ hybridization, we assigned the break point in14qE3 to a 200-kb region devoid of any known gene. We similarly defined

the break point in 15qA1 to a 27-kb region containing involvingADAMTS12.The chromosomal break likely is between exons 2 and 3 ofADAMTS12. Thisgene encodes a disintegrin and metalloproteinase with thrombospondinmotifs, and this product plays crucial roles in both vascularization and cancerprogression and has been implicated in the development of arthritis. TheCBA/CaH-T(14;15)6Ca/J mouse strain likely is a suitable model for furtherexamination of the influences of defective ADAMTS12 in various pathologicprocesses.

Experimental Infection of Mice with Hamster Parvovirus: Evidencefor Interspecies Transmission of Mouse Parvovirus 3

Authors: Christie, Rachel D.1; Marcus, Emily C.1; Wagner, AprilM.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 withclinical signs similar to those induced in hamsters experimentally infectedwith other rodent parvoviruses. Genetically, HaPV is most closely related tomouse parvovirus (MPV), which induces subclinical infection in mice. A novel

MPV strain, MPV3, was detected recently in naturally infected mice, andgenomic sequence analysis indicates that MPV3 is almost identical to HaPV.The goal of the present studies was to examine the infectivity of HaPV inmice. Neonatal and weanling mice of several mouse strains were inoculatedwith HaPV. Tissues, excretions, and sera were harvested at 1, 2, 4, and 8 wkafter inoculation and evaluated by quantitative PCR and serologic assaysspecific for HaPV. Quantitative PCR detected viral DNA quantities that greatlyexceeded the quantity of virus in inocula in multiple tissues of infected mice.Seroconversion to both nonstructural and structural viral proteins wasdetected in most immunocompetent mice 2 or more weeks after inoculationwith 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 at8 wk after inoculation. No clinical signs, gross, or histologic lesions wereobserved. These findings are similar to those observed in mice infected withMPV. These data support the hypothesis that HaPV and MPV3 are likelyvariants of the same viral species, for which the mouse is the natural rodenthost with rare interspecies transmission to the hamster.
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Effects of Murine Norovirus Infection on a Mouse Model of Diet-Induced Obesity and Insulin Resistance

Authors: Paik, Jisun1; Fierce, Yvette1; Drivdahl, Rolf1; Treuting, PiperM.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 UnitedStates, and we currently lack sufficient data to determine whether it shouldbe eliminated. It is generally accepted that the virus does not cause clinicalsymptoms in immuno-competent mice. However, we previously reported thatMNV infection alters the phenotype of a mouse model of bacteria-inducedinflammatory bowel disease in part through its effects on dendritic cells. Thetropism of MNV toward macrophages and dendritic cells makes MNV a

potential intercurrent variable in murine models of macrophage-driveninflammatory diseases, such as obesity, insulin resistance, andatherosclerosis. Therefore, we determined whether MNV infection alteredobesity and insulin resistance phenotypes in C57BL/6 mice, a widely usedmodel 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 subtlechanges in lymphoid tissue. Further studies using other models of metabolicdiseases are needed to provide additional information on the potential rolethis 'subclinical' virus might have on disease progression in mouse models ofinflammatory diseases.

Male CD81 Knockout Genotype Disrupts Mendelian Distribution ofOffspring

Authors: Mordica, Whitney J.1; Gallagher, Ryan J.1; Kennedy, JennaL.1; Chapes, Stephen K.2


CD81 is an integral membrane protein in the tetraspanin superfamily thatserves as an adaptor protein. CD81 is also a maternally imprinted gene that

is found in a regulated cluster of genes on mouse chromosome 7. Amongoffspring produced from heterozygous breeding pairs, CD81null/null mice grewat the same rate as CD81+/+ and CD81+/null mice. Because of an inhibition insperm-egg fusion, CD81null/null female mice are much less fertile than CD81+/+

and CD81+/null mice. However, no published study has detailed the effect ofthe male CD81 genotype on the genotype and sex distribution of offspring.We set up breeding pairs of heterozygotic (C.129-Cd81tm1 N7) female miceand male mice with CD81+/null, CD81+/+, or CD81null/null genotypes. The survival
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and development of CD81+/null, CD81+/+, and CD81null/null offspring weremonitored and compared. Compared with those of heterozygous malebreeders, CD81null/null pups were born at a less-than-expected ratio fromCD81null/null males. Sex distribution did not differ among pups sired byCD81null/null compared with CD81+/null mice. The data suggest that the effect of

the CD81null/null

paternal genotype on offspring is manifested early indevelopment or in utero.

Etiopathogenesis of Mandibulofacial and Maxillofacial Abscesses inMice

Author: Lawson, Gregory W.1


The etiologic agent of mandibulofacial and maxillofacial abscesses in mice isreportedly coagulase-positive Staphylococcus aureus. Although suggested tobe through the oral cavity, the exact route of entry has not beendocumented. Among the clinical cases of mandibulofacial and maxillofacialabscess we report here, each case that was cultured yielded coagulase-positive S. aureus. Histologically, all of the abscesses examined were directlyassociated with intralesional hair shafts, both vibrissae and pelage, that wereintroduced into the submucosa via the maxillary or mandibular molargingival 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, wepropose that mandibulofacial and maxillofacial abscesses are induced by themastication and fragmentation of hair ingested during the barbering process.From the resulting foreign body periodontitis, abscess formation originates atthe maxillary lingual, buccal, or mesial gingival sulci, resulting in infection ofthe maxillary molar tooth roots with swelling or rupture through the skininferior to the eye, or at the mandibular lingual, buccal, and or mesialgingival sulci, resulting in infection of the mandibular molar tooth roots andosteomyelitis with drainage through the skin of the ventral mandible.

Quantitation of Acute Phase Proteins and Protein Electrophoresis inMonitoring the Acute Inflammatory Process in Experimentally andNaturally Infected Mice

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

Serologic screening for infectious disease in sentinel mice from rodentcolonies is expensive and labor-intensive, often involving multiple assays for

several different infectious agents. Previously, we established normalreference ranges for the protein fractions of several laboratory strains ofmice by using a commercially available agarose system of proteinelectrophoresis. In the current study, we address protein fractionation andquantitation of acute phase proteins (APP) in mice experimentally infectedwith Sendai virus or mouse parvovirus. We further investigate thismethodology by using samples from sentinel mice from colonies withendemic infection. All study groups showed significant increases in globulins. Various other protein fractions showed mild variable changes;significant differences were not detected for individual APP. These resultscontrast the significant changes observed in APP and protein electrophoresis

by using the standard methods of inducing inflammatory responses throughinjection of complete Freund adjuvant or LPS. These present data suggestthat although quantitation of individual APP may not be helpful, globulinlevels may reflect infection in laboratory mice and provide a possible adjunctto traditional screening methods.

Lack of Association of a Spontaneous Mutation of the Chrm2 Genewith Behavioral and Physiologic Phenotypic Differences in InbredMice

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 ofleucine for proline at position 266 (P266L) of the CHRM2 protein. BecauseChrm2 codes for the type 2 muscarinic receptor, this mutation couldinfluence physiologic and behavioral phenotypes of mice. Chrm2 mRNA wasnot 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 C797Tpoint mutation in Chrm2 did not differ significantly in muscarinic bindingproperties. Variation across strains was detected in terms of acoustic startle,prepulse inhibition, and the physiologic effects of the muscarinic agonistoxotremorine. However, interstrain differences in these measures did notcorrelate with the presence of the mutation. Although we were unable toassociate a measurable phenotype with the Chrm2 mutation, assessment of
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the mutation on other genetic backgrounds or in the context of other traitsmight reveal differential effects. Therefore, despite our negative findings,evaluation of characteristics that involve muscarinic function should beundertaken with caution when comparing mice with different alleles of theChrm2 gene

Variation in the Gut Microbiota of Laboratory Mice Is Related to BothGenetic and Environmental Factors

Authors: Hufeldt, Majbritt Ravn1; Nielsen, Dennis S.2; Vogensen, FinnKvist2; 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 receivedincreasing attention as a factor in the development of experimentalinflammatory disease in animal models. Because increased variation in theGM might lead to increased variation in disease parameters, determining andreducing GM variation between laboratory animals may provide moreconsistent models. Both genetic and environmental aspects influence thecomposition of the GM and may vary between laboratory animal breedingcenters and within an individual breeding center. This study investigated thevariation in cecal microbiota in 8-wk-old NMRI and C57BL/6 mice by usingdenaturing gradient gel electrophoresis to profile PCR-derived ampliconsfrom 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 micefrom 2 vendors revealed significant differences in the microbial profile,whereas the profiles of C57BL/6Sca mice raised in separate rooms within thesame breeding center were not significantly different. Furthermore, housingin individually ventilated cages did not lead to intercage variation. Theseresults show that denaturing gradient gel electrophoresis is a simple tool thatcan be used to characterize the gut microbiota of mice. Including suchcharacterizations in future quality-control programs may increase thereproducibility 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
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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 problemin laboratory colonies, and diagnosis of MPV infection is complicated by viral

and host factors. The effect of an underlying viral infection on MPV infectionhas not previously been investigated. We assessed the effect of murinenorovirus (MNV) infection, the most prevalent infectious agent in laboratorymice, on MPV shedding, tissue distribution and transmission. Fecal MPVshedding persisted longer in BALB/c mice infected with MNV 1 wk prior toMPV infection than in mice infected with MPV only, but transmission of MPVto soiled-bedding sentinels was not prolonged in coinfected mice. MPV DNAlevels in coinfected BALB/c mice were higher in mesenteric lymph nodes andspleens at 1 and 2 wk after inoculation and in small intestines at 1 wk afterinoculation compared with levels in mice infected with MPV only. In C57BL/6mice, fecal shedding was prolonged, but no difference in soiled bedding

transmission or MPV DNA levels in tissues was detected between singly andcoinfected mice. MPV DNA levels in singly and coinfected SW mice weresimilar. MPV DNA levels were highest in SW, intermediate in BALB/c andlowest in C57BL/6 mice. MPV DNA levels in mesenteric lymph nodes ofBALB/c and SW mice exceeded those in small intestines and feces, whereasthe inverse occurred in C57BL/6 mice. In conclusion, MNV infection increasedthe duration of MPV shedding and increased MPV DNA levels in tissues ofBALB/c mice.

Anomer-Equilibrated Streptozotocin Solution for the Induction of

Experimental Diabetes in Mice (Mus musculus)

Authors: de la Garza-Rodea, Anabel S.1; Knan-Shanzer, Shoshan1; denHartigh, Jan D.2; Verhaegen, Alphons P.L.2; van Bekkum, Dirk W.1


Streptozotocin is widely used to induce diabetes in laboratory animalsthrough multiple low-dose or single high-dose intraperitoneal injections.

HPLC analysis has shown that the composition of the solution may changeconsiderably during the first 2 h after dissolution due to equilibration of the 2anomers ( and ) of streptozotocin. Because of the drug's alleged instabilityin solution, the typical recommendation is to administer streptozotocin within10 min after dissolution. We compared the induction of diabetes in NOD/SCIDmice by injection of a single high dose of freshly made or anomer-equilibrated streptozotocin solution. Solutions were prepared from drycompound containing 85% of the anomer, which is the more toxic of the 2.
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Body weight and nonfasting blood glucose levels were measured weekly for8 wk. Both solutions induced long-term hyperglycemia, but blood glucoselevels and mortality were higher and damage to pancreatic islands morepronounced in the mice receiving freshly prepared solution. A smallproportion 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 atleast 40 d; under those conditions the streptozotocin content decreases by0.1% daily, as determined by HPLC. Anomer-equilibrated streptozotocinsolution has several practical advantages, and we recommend its use asstandard for the induction of experimental diabetes because this practicemay improve reproducibility and comparison of results between differentlaboratories.

Suppurative Adenitis of Preputial Glands Associated withCorynebacterium mastitidis Infection in Mice

Authors: Radaelli, Enrico1; Manarolla, Giovanni2; Pisoni, Giuliano2; Balloi,Annalisa3; Aresu, Luca4; Sparaciari, Paolo5; Maggi, Adriana6; Caniatti,Mario2; Scanziani, Eugenio1


Suppuration of the preputial gland in mice occurs as a septic complication offight wounds around the external genitalia. Currently reported bacterialisolates from these lesions are limited to Staphylococcus aureus, Pasteurella

pneumotropica, and Klebsiella oxytoca. In the context of a pilot experimentaimed at defining the aging phenotype of estrogen receptor 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 tohave preputial gland lesions. In both cases, histopathologic examinationconfirmed severe suppuration and abscesses of the preputial glandsassociated with systemic reactive (secondary) amyloidosis. Both Gramstaining and Bacillus Calmette-Gurin immunohistochemistry highlighted thepresence of numerous bacillary to rod-shaped bacteria within the preputiallesions. Subsequent PCR analysis coupled with denaturing gradient gelelectrophoresis identified Corynebacterium mastitidis in the preputial gland

abscesses. This organism is isolated infrequently from the milk of sheep withsubclinical mastitis and was identified as part of the normal microflora of thehuman ocular surface. No information regarding the epidemiology andpathogenesis ofC. mastitidis infection in laboratory animals is currentlyavailable, and to our knowledge this report is the first description ofC.mastitidis infection in mice.
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Disparities in Ammonia, Temperature, Humidity, and AirborneParticulate Matter between the Micro-and Macroenvironments ofMice 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 theassumption that the environment within the cage closely mirrors the roomenvironment. This study evaluated that premise by examining macro- (room)and microenvironmental (cage) parameters in individually ventilated cageshousing mice with variable amounts of bedding over a period of 17 d without

cage changes. Intracage ammonia levels remained within recommendedhuman guidelines but were higher than room levels, confirming thatmicroisolation caging is efficient at preventing ammonia generated fromanimal waste from escaping into the room. Humidity and temperature withincages were consistently higher than room levels. Particles in the roompredominantly consisted of fine particles (diameter less than 2.5 m),presumably from the ambient atmosphere; some of these particles werefound in the cage microenvironment. In addition, mouse activity within cagesproduced larger particles, and these particles contributed to substantiallyhigher aerosol mass concentrations within the cage. These findingsdemonstrate that, although cage and room environmental parameters differ,

knowledge of room environmental conditions can be used to predict certainconditions within the cage. This association is relevant in that typical animalcare standard operating procedures rely on room measurements, notintracage measurements, which arguably are more important for assessinganimal welfare. Further, location and ambient climate can influence particleconcentrations in the room, and consequently within the animal cage,suggesting local weather patterns and air quality may account for variabilityamong 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: Fernndez, Itziar1; Pea, Arantza1; Del Teso, Nahia1; Prez,Virginia1; Rodrguez-Cuesta, Juan1

Source:Journal of the American Association for Laboratory Animal Science,Volume 49, Number 2, March 2010 , pp. 202-206(5)
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Collection of blood from the submandibular vein allows simple and rapidprocessing of many animals without anesthesia and facilitates rapid recoverywith no signs of pain and discomfort in the mice. Here we compared thesubmandibular vein and retroorbital plexus blood collection methods, todetermine the potential effect of the sampling technique on several clinical

biochemistry parameters in C57BL/6J mice. We found statistically significantdifferences for 8 of the 9 biochemical parameters studied between the 2blood sampling techniques. Compared with samples collected from theretroorbital plexus, blood obtained from the submadibular vein had higherlevels of AST, ALT, protein, albumin, triglycerides, total cholesterol, andcreatinine. Glucose values of retroorbital blood were higher than those fromthe submandibular vein. Urea levels were similar for both samplingtechniques. Our results demonstrate that the technique used to obtain bloodsamples affects parameters commonly used to assess animal health. Werecommend caution when comparing results of biochemical analysis of bloodobtained from the submandibular vein in mice with reference values

obtained by other blood sampling techniques.

Identification of Markers for Imminent Death in Mice used inLongevity 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 reliablypredict imminent death in aged mice. Male and female ICR mice (age, 8 mo)were subcutaneously implanted with an identification chip for remotemeasurement of body temperature. Mice then were weighed and monitoredregularly until spontaneous death occurred or until euthanasia wasadministered for humane reasons. Clinical signs that signaledimplementation of euthanasia included inability to walk, lack of response tomanipulation, large or ulcerated tumors, seizures, and palpable hypothermia.In mice that died spontaneously, gradual weight loss was the most frequentand earliest sign of imminent death. Hypothermia developed during the 2 wk

prior to death. Slow or labored breathing were observed in about half of themice before death. A composite score of temperature weight can be usedto provide an objective benchmark to signal increased observation oreuthanasia of individual mice. Such assessment may allow the collection ofterminal tissue samples without markedly altering longevity data, althoughapplication of this criterion may not be appropriate for all studies oflongevity. Timely euthanasia of mice based on validated markers of imminentdeath can allow implementation of endpoints that alleviate terminal distress
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in aged mice, may not significantly affect longevity data, and can permittimely collection of biologic samples.

Eradication ofHelicobacterspp. by Using Medicated Diet in MiceDeficient in Functional Natural Killer Cells and Complement Factor D

Authors: del Carmen Martino-Cardona, Maria1; Beck, Sarah E.1; Brayton,Cory1; Watson, Julie1


A commercial 4-drug diet has shown promise in eradicating Helicobacterspp.from rodents; however, its effectiveness in immunocompromised mice isunknown. This study evaluated the efficacy of this treatment in eradicating

Helicobacterspp. 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 medicateddiet for 8 wk followed by 4 wk on control diet. Fecal samples were PCR-evaluated for Helicobacterspp. before mice began treatment and then every2 wk thereafter for 12 wk. The same experimental design was repeated foreighteen 9- to 21-wk-old Df-/- mice naturally infected with H. bilis with orwithout H. rodentium. All Df-/- mice and 8- to 21-wk-old Cd1/ mice ceasedshedding Helicobacterspp. after 2 wk of treatment and remained negativethroughout the study. In contrast, the Cd1/ mice that were 24 wk or oldershed Helicobacterspp. for the first 8 wk but tested negative at 10 and 12 wk.

All treated animals had enlarged ceca and gained less weight than controluntreated mice, and 6 of 7 treated Cd1/ male mice developed mild portalfibrosis. These findings show that within 2 wk of treatment, the 4-drug dieteradicated H. hepaticus and H. rodentium from young Cd1/ mice and H.bilis and H. rodentium from Df/ mice, but eradication of establishedinfections in Cd1/ mice required 8 wk of treatment.

Vacuum-Cleaner Noise and Acute Stress Responses in FemaleC57BL/6 Mice (Mus musculus)

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

Katherine3; Francis, Darlene D.4


Audiogenic stress is a well-documented phenomenon in laboratory rodents.Despite the recommendation in the Guide for the Care and Use of LaboratoryAnimals to consider noise a concern in the animal facility, only a small body
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of literature empirically addresses the effects of facility noise on laboratoryrodents, particularly mice. The objective of this study was to determinewhether facility noise generated by a vacuum cleaner induces an acutestress response in a commonly used strain of laboratory mouse undercommon housing conditions. In each of 2 experiments, 10 young adult,

female C57BL/6Cr mice were exposed for 1 h to noise produced by a vacuumcleaner, and 10 control mice were not. In the first experiment, fecal sampleswere collected to measure concentrations of fecal corticosterone metabolitesjust before and 2, 4, 6, 8, 10, 14, 24, and 32 h after noise exposure. In thesecond experiment, stress-sensitive behavioral tests were performed 2 dbefore, immediately after, and 24 h after noise exposure. Physiologic andbehavioral measurements indicated that vacuum cleaner noise did not causean acute stress response in the noise-exposed mice but may have affectedthe diurnal variation of their corticosterone levels. These findings couldcontribute to the development of best practices in noise-control protocols foranimal facilities.

Evaluation of a Commercial Colorimetric Fecal Dipstick Assay for theDetection ofHelicobacter 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


Mice used in biomedical research typically are tested for the presence of

Helicobacterspp., including Helicobacter hepaticus. Here we evaluated theability of a commercially available colorimetric Helicobacterdipstick assay todetect H. hepaticus in experimentally and naturally infected mice, with useof a HelicobacterPCR assay as the 'gold standard' test. None of the fecalsamples from experimentally infected A/JCr mice (n = 12) tested positive forHelicobacterby the colorimetric dipstick test. In naturally infected A/JCr andC57BL/6 mice, 11% (1 of 9) and 30% (3 of 10) of fecal samples, respectively,tested positive for Helicobacterby the colorimetric dipstick assay. In these 3groups ofH. hepaticus-infected mice, statistically fewer mice tested positiveby the colorimetric dipstick test than by PCR. The colorimetric Helicobacterdipstick assay had an overall diagnostic sensitivity of 13%, diagnostic

specificity of 94%, and analytical sensitivity of 108

H. hepaticus cfu/mL. Ascurrently formulated, the colorimetric dipstick assay had high specificity butlacked sensitivity for detecting H. hepaticus infections in 2 strains of micecommonly used in research, thereby limiting its utility as a diagnosticscreening test for H. hepaticus infections in research mice.

Euthanasia by CO2 Inhalation Affects Potassium Levels in Mice


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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 0.85mEq/L or greater in laboratory mice; this concentration has even beenpublished as the upper limit of a 'normal' reference range. However, if bonefide, this potassium concentration would be incompatible with life in allspecies. We investigated conditions frequently encountered in the researchsetting to distinguish artifactual from true hyperkalemia. Variables evaluatedincluded site of collection, time allowed for clot formation before serumseparation, time elapsed between collection and analysis of samplescollected in a serum separator tube, precollection method of anesthesia, and

euthanasia technique. Serum potassium was measured from 75 C57BL/6NTac10-wk-old female mice and divided into at least 5 mice per variable. Animalswere euthanized by exsanguination immediately after terminal CO2 orketamine-xylazine (KX) administration. Mice euthanized with CO2 had highermean serum potassium (7.0 0.5 mEq/L) and range serum potassium (6.0to 8.1 mEq/L) than did KX-treated mice. CO2 inhalation resulted insignificantly 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 followedby exsanguination. These results suggest that antemortem respiratoryacidosis 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 Careafter Total-Body Irradiation in Mice (Mus musculus)

Authors: Moccia, Krinon D.1; Olsen, Cara H.2; Mitchell, JenniferM.1; Landauer, Michael R.1


Concern regarding the potential for radiation exposure from accidents ornuclear and radiologic terrorism is increasing. The purpose of this study wasto determine whether the addition of minimal supportive care consisting ofhydration or nutritional gels could be used to reduce mortality in miceexposed to 60Co -radiation. Male CD2F1 mice received 0, 8.50, or 9.25 Gy60Co at a dose rate of 0.6 Gy/min. These groups were further divided into 3treatment groups thatin addition to pelleted food and waterreceived no
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supportive care, hydration gel, or nutritional gel. Overall survival, meansurvival time, consumption of pelleted food and gel, and body weight wererecorded for 30 d. Radiation caused dose-dependent decreases in overallsurvival, consumption of pelleted food and supplemental gel, and bodyweight. However, at each radiation dose (0, 8.50, 9.25 Gy), the type of

supportive care did not modify overall survival, mean survival time, orchanges in body weight. These results demonstrate that hydration andnutritional gels were not effective methods of supportive care after high-dosetotal body irradiation in mice.

A Spoonful of Sugar Helps the Medicine Go Down: A Novel Techniqueto Improve Oral Gavage in Mice

Authors: Hoggatt, Amber F.1; Hoggatt, Jonathan2; Honerlaw,Meghan3; Pelus, Louis M.2


Oral gavage is a common route of precise oral dosing for studies in rodents.Complications including tracheal administration, esophageal trauma, andaspiration are common and usually related to animal resistance to theprocedure, and the stress induced by oral gavage can be a confoundingvariable in many studies. The taste of sucrose conveys a pacifying andanalgesic effect in newborns, whereas sour solutions can induce the swallow

reflex in humans that are dysphagic. We hypothesized that precoating agavage needle with sucrose or citrate (or both) would pacify mice and inducethem to swallow, reducing the stress and complications associated with thetechnique. To validate this hypothesis, we quantitated time to passage,stress-related behavioral reactions to the procedure, and plasmacorticosterone 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, decreasedobservable stress-related reactions to the procedure, and maintained plasmacorticosterone levels similar to those in ungavaged control mice. Coatingneedles with water, sucrose and citrate, or citrate had no beneficial effects

on these parameters. Our findings describe a novel, validated technique thatmeasurably decreases signs of stress and thereby improves animal welfareduring oral gavage. Furthermore, the use of sucrose may be a valuable toolto refine other minor or nonsurgical procedures in the field of laboratoryanimal research.

Short-Term Storage and Transport at Cold Temperatures of 2-CellMouse Embryos Produced by Cryopreserved Sperm
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A mouse parvovirus (designated MPV1f) was identified in a commerciallaboratory mouse colony in Australia. The infection had not been detected byusing an rNS1 parvovirus ELISA antigen even though the virus wasgenetically similar to other MPV1 variants reported previously. A recombinantbiotinylated protein based on a truncated VP1 protein of the MPV1 strain was

produced and used as antigen for ELISA and Western immunoblots to detectvirus infection and determine the seroprevalence of infection in a colony ofapproximately 45,000 mice. Antibody-positive mice were detected in 8 of 11rooms sampled, indicating that infection was widespread in the facility.Antibody was detected in 16.2% of 1161 sera obtained from 20 strains ofmice. Seroprevalence varied among mouse strains, suggesting geneticvariation in the susceptibility of mice to MPV1 or in their antibody responseto infection, as has been reported previously in experimentally infected mice.Seroprevalence was high in some inbred strains, including DBA/2JArc and therandom-bred strains Hsd:NIH and Arc:Arc(s). Antibody was not detectedinC57BL/6J strains, and BALB/c strains showed low seroprevalence of MPV1f.

Strain- and Age-Associated Variation in Viral Persistence andAntibody Response to Mouse Parvovirus 1 in ExperimentallyInfected 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 andconcurrent antibody response to mouse parvovirus 1 (isolate MPV1f) wasevaluated for 305 d after inoculation in 4 strains of mice. The resultsconfirmed previous reports that mouse strain and age at infection aresignificant factors in viral persistence and antibody development anddetection. Randombred Arc:Arc(s) mice originally bred from CD1 stockinoculated as juveniles (4 wk) or adults (8 wk) developed persistent viralinfection for 152 d after inoculation and an antibody response that persistedfor 295 d. Mice of C57BL/6J background inoculated as juveniles haddetectable viral DNA in large intestinal content and tissues for 24 d afterinoculation 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 resultssuggest probable viral replication in adult C57BL/6J mice but at levels belowthe limits of detection. BALB/cArc mice inoculated as juveniles or adults haddetectable virus DNA in tissues for 108 to 242 d after inoculation, but noantibody was detected. Similarly, BALB/c-Foxn1nu/Arc mice had detectablelevels of viral DNA in tissues for 98 to 131 d but no measurable antibody. The


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difficulty of detecting antibody in mice with a BALB/c background indicatesthey are unsuitable for routine surveillance of MPV1f infection.

Spontaneous Staphylococcus xylosus Infection in Mice Deficient inNADPH Oxidase and Comparison with Other Laboratory Mouse

Strains

Authors: Gozalo, Alfonso S.1; Hoffmann, Victoria J.2; Brinster, LaurenR.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 commoninhabitant of rodent skin. Reports ofS. 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 contributingto the death or need for euthanasia of the animal. Infection with S. xylosuswas the major cause of death or euthanasia in 3 strains of mice deficient inthe production of phagocyte superoxide due to defects in NADPH oxidase.NADPH-oxidase-deficient mice (n = 21) were most susceptible tospontaneous S. xylosus infections. The infections were characterized byabscesses and granulomas in soft tissues, with bacterial migration to internalorgans (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 werelargely confined to eyelids, ocular conjunctiva, and skin and rarely involvedother tissues or organs. Because exhaustive bacterial culture and isolationmay not be performed routinely from mouse abscesses, S. xylosus infectionsmay be underdiagnosed. S. xylosus should be considered in the differentialdiagnosis in laboratory mice with abscesses and other skin lesions. Thisreport expands the range of mouse strains and tissues and organssusceptible to spontaneous S. xylosus infection and compares the pathologyamong various mice strains.

Treatment and Eradication of Murine Fur Mites: I. ToxicologicEvaluation 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)
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Fur mite outbreaks remain a persistent problem in laboratory mousecolonies. All currently published treatment methods are labor-intensive,expensive, or unreliable. During a recent outbreak with Myobia musculi andMyocoptes musculinus in a large colony (approximately 30,000 cages), wedeveloped a feed-based treatment regime in which ivermectin was the active

ingredient. Rodent feed was compounded with 3 different concentrations ofivermectin (12, 24, and 48 ppm) and -irradiated. Postcompounding analysisrevealed loss of ivermectin during manufacturing, but the remaining drugwas stable for at least 6 mo. In an 8-wk toxicity study in a C57BL/6NTacmouse breeding colony, ad-libitum feeding of the 3 diets yielded estimateddoses 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 andataxia and was euthanized. No significant differences between doses wererevealed by CBC, serum chemistry, body weight, or gross necropsy. Plasmadrug concentrations plateaued at a dose-dependent level 7 to 10 d afterinitiation of treatment and decreased to undetectable levels 6 to 9 d after its

discontinuation. Fertility of the P0 generation was unaffected. Pup mortalitywas higher in the 24- and 48-ppm groups, reaching 100% at the higher dose.Animals exposed to ivermectin as neonates had normal weaning weights, butmice receiving 24-ppm feed had lower adult weights. Our results indicatethat using feed containing 12 ppm ivermectin (estimated ingested dose, 1.3mg/kg) was safe in a C57BL/6NTac breeding colony.

Treatment and Eradication of Murine Fur Mites: II. DiagnosticConsiderations

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


Fur mites are a persistent problem in contemporary laboratory mousecolonies. We conducted several studies to evaluate fur mite diagnosticmethodologies and interpretation of results. Retrospective analysis of testresults from sentinel mice exposed to soiled bedding collected from coloniesinfested with Myobia musculi and Myocoptes musculinus revealed the skinscrape test to be more reliable than pelt examination, provided that both the

head and dorsal thoracolumbar regions were sampled. To assess theirdiagnostic accuracy, 3 commercial laboratories were sent positive controlslides containing mites, mite parts, or eggs in sets of slides containingdiagnostic skin scrapings in varying ratios. Laboratory B correctly identifiedthe positive control slide. Laboratory A identified 1 of 3 positive controlslides, whereas laboratory C failed to identify both positive control slidessubmitted. To determine the time required for a mouse to shed its entire haircoat, fur of Crl:CD1(ICR), BALB/cAnNCrl, and Crl:CFW(SW) albino mice was
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dyed black and the presence of dyed fur evaluated monthly for 8 mo. Limiteddyed hair was still present at 8 mo; therefore, finding eggs or egg casingsmany months after treatment cessation does not necessarily imply treatmentfailure. To evaluate the effectiveness of soiled bedding sentinels for detectionof fur mites in a mite-infested colony, we exposed nave mice to varying

amounts (100%, 50%, 25%, 2.5%, and 0%) of soiled bedding in cleanbedding. As little as 2.5% soiled bedding resulted in detection of a positivesentinel within a 2-mo period.

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

Authors:Tavakkol, Zarry1; Samuelson, Derrick2; deLancey Pulcini,Elinor2; Underwood, Robert A.1; Usui, Marcia L.1; Costerton, JWilliam3; James, Garth A.2; Olerud, John E.1; Fleckman, Philip1


Research in cutaneous biology frequently involves models that use micehoused in SPF conditions. Little information is available concerning thespecies of bacteria that normally inhabit the skin of these mice. The aim ofthis study was to characterize the bacterial skin flora of mice housed underSPF conditions. Skin biopsies from C57BL/6 mice under normal and surgicallyprepped conditions were both cultured and analyzed by using DNA extractionand sequencing. The species isolated most commonly from culture were

staphylococci. Coagulase-negative staphylococci were isolated morefrequently than was Staphylococcus aureus. Molecular sequencing yieldedseveral additional organisms not found by culture. Overall, culturing ofisolates yielded 14 species of bacteria, and molecular sequencing identifiedanother 6 species. Investigators conducting cutaneous research in mousemodels should aware of the cutaneous bacterial flora present on these mice.

Noise in a Laboratory Animal Facility from the Human and MousePerspectives

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


The current study was performed to understand the level of sound producedby ventilated racks, animal transfer stations, and construction equipment
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buprenorphine treated mice showing the highest pain index scores, ascompared with nonsurgical controls. Fecal corticosterone metabolite levelsdid not differ significantly between any of the groups or across time. Theparameters used in this study did not indicate that administration of theseanalgesic regimens improved recovery as compared with that of saline-

treated mice. Care should be taken when using visual assessment scores toevaluate pain in mice, given that analgesics may have side effects thatinadvertently elevate the score.

Treatment and Eradication of Murine Fur Mites: III. Treatment of aLarge 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 furmites in mice and describe its use to eradicate mites in vivaria holdingapproximately 30,000 cages. C57BL/6NCrl mice infested with Myobia musculiand 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 contactsentinels, remained free of fur mites for as long as 21 wk after treatment. Noadverse effects were observed. Subsequently, facility-wide treatment wasimplemented in an attempt to eradicate fur mites from 3 vivaria housing

approximately 120,000 mice. Medicated feed was provided for 8 wk toensure that all cages and mice were treated. A single investigative groupreported adverse effects in their colony 4 wk after treatment was initiated;mortality was attributed to ivermectin toxicity after an intracranial injectionat 1 d of age. Nave pups were unaffected. No other adverse effects werenoted. Approximately 14,500 skin scrape samples were evaluated during the12-mo posttreatment surveillance period. All samples were negative formites. To our knowledge, this is the first report of successful eradication offur 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, Joo P.L.2; Hutchinson, EricK.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)
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and sterile surgical gloves were used for each procedure. In this study, weobserved that the modified aseptic technique using 70% isopropyl alcoholsoaks pre- vented aerobic bacterial contamination of instruments and glovesfor as many as 5 mice.

2010 mouse journal abstracts

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