PLASMA ELECTROPHORETIC PROFILES AND HEMOGLOBIN BINDING PROTEIN REFERENCE INTERVALS IN THE EASTERN BOX TURTLE (TERRAPENE CAROLINA CAROLINA) AND INFLUENCES OF AGE, SEX, SEASON, AND LOCATION

Abstract

Evaluation of plasma electrophoretic profiles and acute phase protein concentrations may play a valuable role in health assessment of reptiles; however, little is known about reference intervals in free-ranging eastern box turtles (Terrapene carolina carolina). The purpose of this study was to establish reference intervals of protein electrophoretic profiles and hemoglobin binding protein ([HBP] as determined by a haptoglobin assay) in free-ranging eastern box turtles and to assess any possible correlations between varying age class (adults vs. juvenile), sex (male, female, or unknown), season (spring, summer, or fall), or location (Tennessee vs. Illinois). Blood samples were obtained from 324 eastern box turtles from 2010 to 2012 at three sites in Illinois and one site in Tennessee, USA. Significant differences were observed with total protein (sex, season, state, Illinois location), albumin (age class, season, state, Illinois location), α-1 globulins (sex, season, Illinois location), α-2 globulins (sex, season, state, Illinois location), β globulins (age class, sex, season, state, Illinois location), γ globulins (sex, season state, Illinois location), and hemoglobin binding protein (age class, sex, state, Illinois location). The use of electrophoretic profiles and acute phase proteins is a relatively new concept in reptilian medicine, and this study allowed for establishment of references intervals in the eastern box turtle and emphasized differences that occured based on age, sex, season, and location. Future research in this area can now build on these data to determine changes in population health over time or alterations due to specific environmental or disease threats.

Erythrocyte sedimentation rate and hemoglobin-binding protein in free-living box turtles (Terrapene spp.)

“…In T. ornata, HBP concentrations were linearly quantifiable and precise, with an inter-assay CV of 4.23% and an intra-assay CV of 3.82%. These values are comparable to those previously reported for T. carolina, (6.6% intra-assay CV) [23] and to those published by the Winpette kit manufacturer (4.1-5.7% inter-assay CV, 5.3-6.3% intra-assay CV). The limit of detection for HBP in T. ornata (0.01 mg/mL) was higher than that reported by the manufacturer (0.005 mg/mL), but lower than the cutoff for T. carolina (0.08 mg/mL) [23].…”

“…Assessing wildlife health status can be challenging due to poor antemortem recognition of disease, limited understanding of pathogen epidemiology, and a lack of validated diagnostic tests [16,17]. Exacerbating these issues, wild reptiles display significant physiologic variation in clinical pathology values based on age, sex, season, and reproductive state [18][19][20][21][22][23][24][25]. For these reasons, routine diagnostic tests such as hematology and plasma biochemistries frequently have poor discriminatory power for identifying unhealthy reptiles [25].…”

“…HBP quantification uses commercially available colorimetric kits that detect the binding of HBP to hemoglobin, and previous studies suggest that the results are reliable across species [1,43,44]. HBP concentrations have been reported in free-living T. carolina [23], but this diagnostic tool has not yet been explored in T. ornata.…”

“…Fourth, to investigate the association between ESR, HBP, and health status. Based on previous studies, we hypothesized that there would be good agreement between the three ESR measurement methods [53][54][55], ESR would be negatively associated with packed cell volume (PCV) [45,46], and both ESR and HBP would elevate in adults [10,23,49,[56][57][58][59], females [23,[59][60][61][62][63][64], and turtles with clinical signs of illness or injury [15,48,[65][66][67].…”

Venous blood gas in free-living eastern box turtles (Terrapene carolina carolina) and effects of physiologic, demographic and environmental factors

“…However, the dramatic physiologic changes associated with growth, reproduction and brumation in reptiles translate to extreme variability in bloodwork parameters based on season, sex, age class and reproductive status (e.g. Anderson et al , 1997 ; Flower et al , 2014 ; Rose and Allender, 2011 ; Tamukai et al , 2011 ). It is therefore important to define the range of expected normal values in free-living populations prior to applying blood gas panels for clinical health assessment.…”