title: Prevalence and diagnosis of hemotrophic mycoplasma infection in research sheep and its effects on hematology variables and erythrocyte membrane fragility.
abstract: Hemotrophic mycoplasma (hemoplasma) infection in research sheep can confound experimental results and contribute to morbidity and mortality. Prevalence and clinicopathologic studies historically relied on blood-smear diagnosis, but systematic studies using current molecular techniques are warranted. Here we sought to report the prevalence of subclinical infection in our study population, compare diagnostic sensitivity and specificity between blood smears and a PCR assay, and determine the effects of infection on CBC variables and erythrocyte membrane fragility. We collected whole-blood samples from 111 convenience-sampled research sheep. All samples were tested for hemoplasmas by using a PCR assay, blood smears were evaluated for visual presence of hemoplasmas, and CBC and osmotic fragility assays were performed. Subclinical prevalence, according to PCR diagnosis, was 14.1% (14 of 99) in our study population. Relative to the PCR assay, blood-smear diagnosis was 8.3% sensitive and 100% specific for hemoplasma detection. Subclinical infection was associated with changes in MCV, MCHC, RBC distribution width, and absolute monocyte count. Acute infection was associated with changes in RBC mass, Hgb concentration, MCV, MCH, MCHC, and absolute lymphocyte and monocyte counts. Acute infection was associated with increased mean erythrocyte fragility compared with that in uninfected control and treated sheep. We demonstrated that hemoplasma infection is common in our study population, blood-smear evaluation is insensitive at detecting infection, and infection is associated with changes in CBC variables and increased erythrocyte membrane fragility. These findings raise concerns regarding the suitability of hemoplasma-infected sheep for biomedical research.
authors.affiliation: Unit for Laboratory Animal Medicine, University of Michigan Medical School, Ann Arbor, Michigan, USA. jahampeldvm@gmail.com.
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Hampel
authors.affiliation: College of Veterinary Medicine, College of Veterinary Medicine, Michigan State University, Lansing, Michigan, USA.
authors.forename: Samantha N
authors.initials: SN
authors.lastname: Spath
authors.affiliation: Unit for Laboratory Animal Medicine, University of Michigan Medical School, Ann Arbor, Michigan, USA.
authors.forename: Ingrid L
authors.initials: IL
authors.lastname: Bergin
authors.affiliation: Diagnostic Center for Population and Animal Health, College of Veterinary Medicine, Michigan State University, Lansing, Michigan, USA.
authors.forename: Ailam
authors.initials: A
authors.lastname: Lim
authors.affiliation: Diagnostic Center for Population and Animal Health, College of Veterinary Medicine, Michigan State University, Lansing, Michigan, USA.
authors.forename: Steven R
authors.initials: SR
authors.lastname: Bolin
authors.affiliation: Unit for Laboratory Animal Medicine, University of Michigan Medical School, Ann Arbor, Michigan, USA.
authors.forename: Melissa C
authors.initials: MC
authors.lastname: Dyson
mesh_terms: D000818:Animals; D000830:Animals, Laboratory; D001399:Azure Stains; D002462:Cell Membrane / Q000473:pathology; D017931:DNA Primers / Q000235:genetics; D004912:Erythrocytes / Q000382:microbiology / Q000473:pathology; D009175:Mycoplasma Infections / Q000097:blood / Q000175:diagnosis / Q000453:epidemiology / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D015995:Prevalence; D012680:Sensitivity and Specificity; D012756:Sheep; D012757:Sheep Diseases / Q000097:blood / Q000175:diagnosis* / Q000453:epidemiology*
chemical_list: D001399:Azure Stains; D017931:DNA Primers