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Experimental studies suggest involvement of trimethylamine N-oxide (TMAO) in the aetiology of cardiometabolic diseases and chronic kidney disease (CKD), in part via metabolism of ingested food. Using a comparative biomimetic approach, we have investigated circulating levels of the gut metabolites betaine, choline, and TMAO in human CKD, across animal species as well as during hibernation in two animal species. Betaine, choline, and TMAO levels were associated with renal function in humans and differed significantly across animal species. Free-ranging brown bears showed a distinct regulation pattern with an increase in betaine (422%) and choline (18%) levels during hibernation, but exhibited undetectable levels of TMAO. Free-ranging brown bears had higher betaine, lower choline, and undetectable TMAO levels compared to captive brown bears. Endogenously produced betaine may protect bears and garden dormice during the vulnerable hibernating period. Carnivorous eating habits are linked to TMAO levels in the animal kingdom. Captivity may alter the microbiota and cause a subsequent increase of TMAO production. Since free-ranging bears seems to turn on a metabolic switch that shunts choline to generate betaine instead of TMAO, characterisation and understanding of such an adaptive switch could hold clues for novel treatment options in burden of lifestyle diseases, such as CKD.

Ecological nutrition aims to unravel the extensive web of nutritional links that drive animals in their interactions with their habitat. Metabolomic profiling of species could increase the knowledge of their ecology, as well as allowing us to better understand the interactions of environment, including distribution, availability of food and fitness of the species, among others. Metabolomic profile is affected by several causes such as nutrition, physiological status, metabolism and species. Thus, the main aim of this work will be to provide, for the first time, values of the nutritional metabolome profile in marine species, as well as to study the differences in the metabolomic profile due to the classification group. To achieve this, blood samples from marine animals (n = 55) were used in this experiment, including Gentoo penguin (Pygoscelis papua), Patagonian sea lion (Otaria flavescens), Harbour seal (Phoca vitulina) and Bottlenose dolphin (Tursiops truncatus). Blood samples were analysed to determine the nutritional metabolites, specifically non‐esterified fatty acid (NEFA), glucose, triglyceride, uric acid, cholesterol, albumin, total protein and inorganic phosphorous (iP). Pygoscelis papua exhibited the most statistically significant differences compared to the other groups, showing higher levels of plasma glucose, triglycerides, uric acid, and cholesterol, and lower levels of albumin, total protein, urea, and iP relative to the average values (simple means) observed in the other species. Also, it can be concluded that nutritional metabolome of Tursiops truncatus is different from that of Otaria flavescens, due to the differences in urea and albumin. These results show the potential of metabolomics to elucidate the relationship between metabolism and external factors in species. However, more molecular ecology studies are necessary to deepen the study of the interrelationships between the environment and metabolism, with the aim of including this tool in specific programmes for conservation and biological study. This study profiles the nutritional metabolomes of four marine species. Pygoscelis papua showed higher glucose, triglycerides, uric acid, and cholesterol, but lower albumin, total protein, urea, and inorganic phosphorus compared to others. Distinct metabolomic differences were also observed between T. truncatus and O. flavescens in urea and albumin levels. These findings highlight the need for further research on environmental impacts on metabolism for conservation and ecological studies.

To date, there is limited knowledge of the effects that abnormal sea surface temperature (SST) can have on the physiology of neonate pinnipeds. However, maternal nutritional deficiencies driven by alimentary restrictions would expectedly impact pinniped development and fitness, as an adequate supply of nutrients is essential for growth and proper functioning of all body systems, including red blood cell synthesis and clearance. Here, we investigated red blood cell morphology of California sea lion (CSL) pups from the San Benito Archipelago born during the 2014 and 2015 anomalously high SST events recorded in the northeastern Pacific Ocean. We examined whether atypical erythrocyte morphologies were more common in 2015, when the high SST event was more pronounced, and whether the stable isotope signature of pup fur, as an indicator of maternal feeding strategies, accounted for the number of atypical cells. Various atypical erythrocyte morphologies were more prevalent and more abundant than reference values. Evidence of iron deficiency was found in both years, and only pups born in 2014 showed evidence of active erythropoiesis. Microcytes and reticulocytes were more common in pups with higher isotopic δ 13C and lower δ 15N values, suggesting a probable relationship between maternal feeding strategies and the effect of climatic anomalies on red blood cell physiology of their pups. As developing pinnipeds require increased oxygen storage capacity for diving and foraging, the presence of atypical erythrocytes could be relevant to CSL pup fitness if the underlying cause is not reverted. This study is a first step to explore the effects that climatic alterations in the marine environment can have on the blood physiology of developing individuals.

The International Species Information System has set forth an extensive database of reference intervals for zoologic species, allowing veterinarians and game park officials to distinguish normal health parameters from underlying disease processes in captive wildlife. However, several recent studies comparing reference values from captive and free-ranging animals have found significant variation between populations, necessitating the development of separate reference intervals in free-ranging wildlife to aid in the interpretation of health data. Thus, this study characterizes reference intervals for six biochemical analytes, eleven hematologic or immune parameters, and three hormones using samples from 219 free-ranging African lions (Panthera leo) captured in Kruger National Park, South Africa. Using the original sample population, exclusion criteria based on physical examination were applied to yield a final reference population of 52 clinically normal lions. Reference intervals were then generated via 90% confidence intervals on log-transformed data using parametric bootstrapping techniques. In addition to the generation of reference intervals, linear mixed-effect models and generalized linear mixed-effect models were used to model associations of each focal parameter with the following independent variables: age, sex, and body condition score. Age and sex were statistically significant drivers for changes in hepatic enzymes, renal values, hematologic parameters, and leptin, a hormone related to body fat stores. Body condition was positively correlated with changes in monocyte counts. Given the large variation in reference values taken from captive versus free-ranging lions, it is our hope that this study will serve as a baseline for future clinical evaluations and biomedical research targeting free-ranging African lions.

A prospective study to assess changes in selected plasma biochemistry and electrolyte values, plasma insulin and aldosterone concentrations, and electrocardiography (ECG) was performed on eight female captive tigers (Panthera tigris) and three lions (Panthera leo) undergoing general anesthesia for elective laparoscopic ovariectomy. Each animal was sedated with medetomidine (18–25 μg/kg) and midazolam (0.06–0.1 mg/kg) intramuscularly, and anesthesia was induced with ketamine (1.9–3.5 mg/kg) intramuscularly and maintained with isoflurane. Venous blood samples were collected and analyzed for plasma biochemistry parameters and insulin and aldosterone concentrations. An ECG was recorded at the time of each blood sample collection. Mean plasma potassium, glucose, phosphorus, and aldosterone concentrations increased during anesthesia (P ≤ 0.05). One tiger developed hyperkalemia (6.5 mmol/L) 2.5 hr after anesthetic induction. Plasma insulin concentrations were initially below the low end of the domestic cat reference interval (72–583 pmol/L), but mean insulin concentration increased (P ≤ 0.05) over time compared with the baseline values. Three tigers and two lions had ECG changes that were representative of myocardial hypoxemia. Based on these results, continuous monitoring of clinical and biochemical alterations during general anesthesia in large nondomestic felids is warranted, and consideration should be given to reversal of medetomidine in these animals should significant changes in electrolytes or ECG occur.

Domoic acid toxicosis (DAT) in California sea lions (Zalophus californianus) is caused by exposure to the marine biotoxin domoic acid and has been linked to massive stranding events and mortality. Diagnosis is based on clinical signs in addition to the presence of domoic acid in body fluids. Chronic DAT further is characterized by reoccurring seizures progressing to status epilepticus. Diagnosis of chronic DAT is often slow and problematic, and minimally invasive tests for DAT have been the focus of numerous recent biomarker studies. The goal of this study was to retrospectively profile plasma proteins in a population of sea lions with chronic DAT and those without DAT using two dimensional gel electrophoresis to discover whether individual, multiple, or combinations of protein and clinical data could be utilized to identify sea lions with DAT. Using a training set of 32 sea lion sera, 20 proteins and their isoforms were identified that were significantly different between the two groups (p<0.05). Interestingly, 11 apolipoprotein E (ApoE) charge forms were decreased in DAT samples, indicating that ApoE charge form distributions may be important in the progression of DAT. In order to develop a classifier of chronic DAT, an independent blinded test set of 20 sea lions, seven with chronic DAT, was used to validate models utilizing ApoE charge forms and eosinophil counts. The resulting support vector machine had high sensitivity (85.7% with 92.3% negative predictive value) and high specificity (92.3% with 85.7% positive predictive value). These results suggest that ApoE and eosinophil counts along with machine learning can perform as a robust and accurate tool to diagnose chronic DAT. Although this analysis is specifically focused on blood biomarkers and routine clinical data, the results demonstrate promise for future studies combining additional variables in multidimensional space to create robust classifiers.

This study establishes a relationship between positive canine heartworm (Dirofilaria immitis) test results frequently observed in California sea lions (Zalophus californianus) and infection with the filarid nematode Acanthocheilonema odendhali. Four commercially available canine heartworm antigen tests were evaluated for cross-reaction with A. odendhali in California sea lions. Sera were tested from fifteen California sea lions with A. odendhali-associated microfilaremia, confirmed by blood smear, and with no evidence of D. immitis infection at necropsy. Ninety-five percent of tests were falsely positive for D. immitis. This study also determined that the prevalence of A. odendhali infection in stranded California sea lions from central California is approximately 23% by comparing the number of findings of mircofilaremia to the total number of California sea lions sampled at The Marine Mammal Center between 2005 and 2011, inclusive. Acanthocheilonema odenhali microfilaremia in California sea lions is likely to cross-react with canine heartworm antigen tests, and clinicians should interpret results with caution.

Otariids, like other wild mammals, contend with a wide variety of energetic demands across seasons. However, due to the cryptic behaviors of this marine group, few studies have been able to examine longitudinal energetic costs or the potential impact of these costs on seasonal or annual prey requirements. Here we evaluated the changes in energy demand and intake of female California sea lions (Zalophus californianus) during reproductive ( \\documentclass{aastex} \\usepackage{amsbsy} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{bm} \\usepackage{mathrsfs} \\usepackage{pifont} \\usepackage{stmaryrd} \\usepackage{textcomp} \\usepackage{portland,xspace} \\usepackage{amsmath,amsxtra} \\usepackage[OT2,OT1]{fontenc} \\newcommand\\cyr{ \\renewcommand\\rmdefault{wncyr} \\renewcommand\\sfdefault{wncyss} \\renewcommand\\encodingdefault{OT2} \\normalfont \\selectfont} \\DeclareTextFontCommand{\\textcyr}{\\cyr} \\pagestyle{empty} \\DeclareMathSizes{10}{9}{7}{6} \\begin{document} \\landscape $n=2$ \\end{document} sea lions) and nonreproductive ( \\documentclass{aastex} \\usepackage{amsbsy} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{bm} \\usepackage{mathrsfs} \\usepackage{pifont} \\usepackage{stmaryrd} \\usepackage{textcomp} \\usepackage{portland,xspace} \\usepackage{amsmath,amsxtra} \\usepackage[OT2,OT1]{fontenc} \\newcommand\\cyr{ \\renewcommand\\rmdefault{wncyr} \\renewcommand\\sfdefault{wncyss} \\renewcommand\\encodingdefault{OT2} \\normalfont \\selectfont} \\DeclareTextFontCommand{\\textcyr}{\\cyr} \\pagestyle{empty} \\DeclareMathSizes{10}{9}{7}{6} \\begin{document} \\landscape $n=3$ \\end{document} ) periods. Monthly measurements included resting metabolic rate, blood hormone levels, body condition (blubber thickness and body mass), and caloric intake for adult sea lions throughout molting, late pregnancy, lactation, and postweaning. We found that maintenance energy demands decreased from 32.0 to 23.1 MJ d−1before pupping, remaining stable at \\documentclass{aastex} \\usepackage{amsbsy} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{bm} \\usepackage{mathrsfs} \\usepackage{pifont} \\usepackage{stmaryrd} \\usepackage{textcomp} \\usepackage{portland,xspace} \\usepackage{amsmath,amsxtra} \\usepackage[OT2,OT1]{fontenc} \\newcommand\\cyr{ \\renewcommand\\rmdefault{wncyr} \\renewcommand\\sfdefault{wncyss} \\renewcommand\\encodingdefault{OT2} \\normalfont \\selectfont} \\DeclareTextFontCommand{\\textcyr}{\\cyr} \\pagestyle{empty} \\DeclareMathSizes{10}{9}{7}{6} \\begin{document} \\landscape $19.4\\pm 0.6$ \\end{document} MJ d−1during lactation and postweaning. Energy intake rates to meet these demands showed marked changes with activity level and the reproductive cycle, reaching a peak intake of 3.6 times baseline levels during lactation. Translating this into prey demands, we find that 20,000 reproductively active females on San Nicolas Island rookeries would maximally require 4,950 metric tons of Pacific whiting during a month of the breeding season. This localized impact is reduced significantly with postbreeding dispersal and demonstrates the importance of considering spatial and temporal factors driving the energetic requirements of predators when designing marine protected areas.

Levels of zinc (Zn), copper (Cu), potassium (K), calcium (Ca), magnesium (Mg), and phosphorus (P) in plasma of Otaria flavescens females ( n  = 29) were evaluated. Reference intervals were established for each element, being the first report for this species.

Blood chemistry and hematologic reference ranges are useful for population health assessment and establishing a baseline for future comparisons in the event of ecosystem changes due to natural or anthropogenic factors. The objectives of this study were to determine if there was any population spatial structure for blood variables of Steller sea lion (Eumetopias jubatus), an established sentinel species, and to report reference ranges for appropriate populations using standardized analyses. In addition to comparing reference ranges between populations with contrasting abundance trends, data were examined for evidence of disease or nutritional stress. From 1998 to 2011, blood samples were collected from 1,231 pups captured on 37 rookeries across their Alaskan range. Reference ranges are reported separately for the western and eastern distinct population segments (DPS) of Steller sea lion after cluster analysis and discriminant function analysis (DFA) supported underlying stock structure. Variables with greater loading scores for the DFA (creatinine, total protein, calcium, albumin, cholesterol, and alkaline phosphatase) also were greater for sea lions from the endangered western DPS, supporting previous studies that indicated pup condition in the west was not compromised during the first month postpartum. Differences between population segments were likely a result of ecological, physiological, or age related differences.