Reference Intervals in Combined Veterinary Clinical Examinations of Male Black-Spotted Pond Frogs (Pelophylax nigromaculatus)

Abstract

In conservation physiology, analyzing the physiological response of an organism to understand its ability to adapt to environmental changes is a key technique in establishing a successful conservation strategy. Veterinary clinical examinations determine the physiological condition of animals accurately and safely, and this examination is synergistic when combined. The accuracy and safety of a clinical examination makes it advantageous for use in amphibians with high species diversity and numerous endangered species. However, it is necessary to establish a reference interval (RI) for precise interpretations and identification of animals with abnormalities through individual unit testing. We have established RIs for the immunity, serum components, bone mineral density (BMD), and body composition of black-spotted pond frogs (Pelophylax nigromaculatus). Black-spotted pond frogs are a common species and are widely distributed in East Asia, with suitable characteristics for environmental monitoring. Serum was extracted from 151 male frogs to establish the RI for bacterial killing ability in order to represent immunity. We also used the serum to establish an RI of ten additional serum components to determine the nutritional status, organ function status, body osmotic pressure, and homeostasis conditions. The BMD and three body composition measures for diagnosing food intake status and nutritional condition were measured using dual energy X-ray absorptiometry. The RI was recorded as the mean ± standard deviation, median, first (25%) to third (75%) quantile range, 95% confidence interval of the mean and median, and the 95% percentile (2.5%–97.5% range) of all components. The use of combined clinical veterinary examinations aids our understanding of the physiological conditions of an individual according to biotic and abiotic factors on a complex spatiotemporal scale in an ecosystem.

Examination of Physiological and Morphological Differences between Farm-Bred and Wild Black-Spotted Pond Frogs (Pelophylax nigromaculatus)

“…To diagnose various stressors or health conditions of individuals using this analysis, physiological reference intervals (RIs) according to various environmental conditions such as season, sex, and life history are required. Although we have previously established RIs of body composition and serum components for male black-spotted pond frogs [ 69 ], these RIs were not used in the present study because there was a comparison group and there were no RIs for females. However, the establishment of RIs is sufficiently meaningful in that a comparison group is not necessary, and because it can be used for diagnosis on an individual unit [ 26 , 70 ].…”

“…The factors we measure are those that can change more within a specific period of one generation than across several generations. [ 48 , 69 ]. Later, studies of factors that occur across generations, such as changes in the gene pool, may be necessary.…”

Framework for multi-stressor physiological response evaluation in amphibian risk assessment and conservation

“…Neutrophil to lymphocyte ratios were also a good indication of environmental stressors and were associated with total dissolved solid levels in aquatic habitats that impacted growth and development (Ruso et al, 2021). Combinations of physiological indices are also informative to link endpoints with individual condition (e.g., Park et al, 2021).…”

Characteristics of the interferon regulatory factor IRF5 and its stimulation expression by Elizabethkingia miricola Infection in black-spotted frog (Pelophylax nigromaculatus)

“…The black-spotted frog, Pelophylax nigromaculatus, belongs to Pelophylax, Ranidae, extensively distributed in east Asia. 17 Since the success of rearing, it on an artificial diet, black-spotted frog has been widely farmed in China due to its growing fast, delicious, and nutritious, taking on great economic significance. However, a considerable number of black-spotted frogs in farms in Hunan Province in southcentral China have been subjected to an epidemic meningitis-like disease (i.e., "frog torticollis" or "frog cataract") since 2016.…”