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• MWCNT-poly(STY-DVB) composite was synthesized as magnetic solid-phase support. • A novel composite-based approach was used to extract different catecholamines. • A new LC-MS method for determining five catecholamines was developed and validated. • Red deer urine and hair samples were successfully analyzed using this methodology. A novel analytical methodology for the extraction and determination of catecholamines (dopamine, epinephrine and norepinephrine) and their metabolites DL-3,4-dihydroxyphenyl glycol and DL-3,4-dihydroxymandelic acid by LC-MS is developed and validated for its application to human and animal urine and hair samples. The method is based on the preliminary extraction of the analytes by a magnetic multi-walled carbon nanotube poly(styrene-co-divinylbenzene) composite. This is followed by a <9 min chromatographic separation of the target compounds in an Onyx Monolithic C 18 column using a mixture of 0.01% (v/v) heptafluorobutyric acid in water and methanol at 500 µL min−1 flow rate. Detection limits within range from 0.055 to 0.093 µg mL−1, and precision values of the response and retention times of analytes were >90%. Accuracy values comprised the range 79.5–109.5% when the analytes were extracted from deer urine samples using the selected MMWCNT-poly(STY-DVB) sorbent. This methodology was applied to real red deer urine and hair samples, and concentrations within range from 0.05 to 0.5 µg mL−1 for norepinephrine and from 1.0 to 44.5 µg mL−1 for its metabolite 3,4-dihydroxyphenyl glycol were calculated. Analyses of red deer hair resulted in high amounts of 3,4-dihydroxyphenyl glycol (0.9–266.9 µg mL−1). [ABSTRACT FROM AUTHOR]

Developing baseline concentrations of serotonin in healthy white-tailed deer will allow for the development of a biomarker using non-invasive sample tissues in sick animals, for example, non-clinical cases of chronic wasting disease. It will also allow some further insight into whether the use of antibiotics as growth promoters (AGP), such as chlortetracycline, is affecting serotonin concentrations in white-tailed deer. Florfenicol and tulathromycin impacts on serotonin concentration changes were also investigated. An analytical method for the detection and confirmation of serotonin, 5-hydroxytryptamine (5-HT), in white-tailed deer tissues was developed and validated. Serum and urine samples were extracted with acetonitrile. Liquid chromatography separation was attained on a Phenomenex C 18 column with a Security Guard ULTRA guard column with gradient elution using a mobile phase of 0.1% formic acid in water and 0.1% formic acid in acetonitrile. This methodology was applied to baseline (control), chlortetracycline (CTC) treated, florfenicol treated and tulathromycin treated white-tailed deer serum and urine samples., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Chronic wasting disease (CWD) is a prion disease of captive and free-ranging deer (Odocoileus spp), elk (Cervus elaphus nelsonii) and moose (Alces alces shirasi). Unlike in most other prion diseases, in CWD prions are shed in urine and feces, which most likely contributes to the horizontal transmission within and between cervid species. To date, CWD ante-mortem diagnosis is only possible by immunohistochemical detection of protease resistant prion protein (PrP (Sc) ) in tonsil or recto-anal mucosa-associated lymphoid tissue (RAMALT) biopsies, which requires anesthesia of animals. We report on detection of CWD prions in urine collected from pre-symptomatic deer and in fecal extracts by using real time quaking-induced conversion (RT-QuIC). This assay can be useful for non-invasive pre-symptomatic diagnosis and surveillance of CWD.

Innate predator recognition confers a survival advantage to prey animals. We investigate whether giant pandas exhibit innate predator recognition. We analyzed behavioral responses of 56 naive adult captive giant pandas (Ailuropoda melanoleuca), to urine from predators and non-predators and water control. Giant pandas performed more chemosensory investigation and displayed flehmen behaviors more frequently in response to predator urine compared to both non-predator urine and water control. Subjects also displayed certain defensive behaviors, as indicated by vigilance, and in certain cases, fleeing behaviors. Our results suggest that there is an innate component to predator recognition in captive giant pandas, although such recognition was only slight to moderate. These results have implications that may be applicable to the conservation and reintroduction of this endangered species.

A SYTO9 real-time polymerase chain reaction assay for detection of pathogenic Leptospira spp. based on amplification of DNA gyrase subunit B (gyrB) gene has been optimized and evaluated for sensitivity and specificity on kidney and urine samples of New Zealand farmed deer. The detection limit was 10(3) cells/ml (2-10 copies/reaction). Comparison of the assay on deer kidneys (n = 268) with culture as the gold standard revealed a sensitivity and specificity of 85% and 99.2%, respectively. For deer urine (n = 113), the assay was compared with known inoculated samples and revealed a sensitivity and specificity of 96.7% and 100%, respectively. The assay was applied for quantifying pathogenic leptospires shed naturally in deer urine and revealed a detectable concentration of 3.7 × 10(3) to 1.7 × 10(6) cells/ml. To assess the assay's capability for identifying pathogenic Leptospira spp., 14 field isolates of L. borgpetersenii serovar Hardjo-bovis and L. interrogans serovar Pomona were amplified for polymerase chain reaction (PCR) product, purified, and sequenced. When compared with the National Center for Biotechnology Information database, sequence data matched with L. borgpetersenii serovar Hardjo-bovis in 13 samples and L. interrogans serovar Pomona in 1 sample, which was consistent with the microscopic agglutination test (MAT). Sequence analysis of purified PCR product amplified directly from kidney and urine samples also yielded serovar-comparable MAT results. Results suggest that the assay is rapid, sensitive, and specific for detection of pathogenic leptospires in deer clinical samples. The developed assay can also be used for estimating the concentration of leptospires and identifying Leptospira spp. in combination with DNA sequencing.

Wild and domestic ungulates modify their behaviour in the presence of olfactory and visual cues of predators but investigations have not exposed a domestic species to a series of cues representing various predators and other ungulate herbivores. We used wolf (Canis lupus), mountain lion (Puma concolor), and mule deer (Odocoileus hemionus) stimuli (olfactory and visual), and a control (no stimuli) to experimentally test for differences in behaviour of cattle (Bos taurus) raised in Arizona. We measured (1) vigilance, (2) foraging rates, (3) giving up density (GUD) of high quality foods and (4) time spent in high quality forage locations in response to location of stimuli treatments. In general, we found a consistent pattern in that wolf and deer treatments caused disparate results in all 4 response variables. Wolf stimuli significantly increased cattle vigilance and decreased cattle foraging rates; conversely, deer stimuli significantly increased cattle foraging rate and increased cattle use of high quality forage areas containing stimuli. Mountain lion stimuli did not significantly impact any of the 4 response variables. Our findings suggest that domestic herbivores react to predatory stimuli, can differentiate between stimuli representing two predatory species, and suggest that cattle may reduce antipredatory behaviour when near heterospecifics.

The responses of adult female blacklegged ticks, Ixodes scapularis Say, to urine from white-tailed deer, Odocoileus virginianus (Zimmermann), belonging to 4 reproductive categories (doe in estrous, doe out of season, reproductive [dominant] buck, young buck) and to a mixture of urine from nondominant bucks in rut, young bucks out of rut, and nonestrous does were studied in laboratory behavioral bioassays. In high humidity (approximately 95% RH) in a glove box there were no statistically significant arrestment responses to any of the 5 types of urine, but an avoidance response was observed to urine from dominant reproductive bucks. When ticks were tested at approximately 50% RH, with samples of all 5 types of urine in the glove box, significant arrestant responses by the ticks were elicited by urine from does in estrous and by dominant reproductive bucks. When tested without other types of urine in the glove box, the urine mixture elicited an arrestant response at 50% RH. In some circumstances, adult I. scapularis may possibly use deer urine as a chemical cue in selecting host-ambush sites.

In laboratory bioassays under conditions of high humidity, host-seeking female black-legged ticks, Ixodes scapularis Say, avoided urine of reproductively active male and nonestrous female white-tailed deer, Odocoileus virginianus (Zimmermann), the principal host species of the adult stage. At 50% RH, female I. scapularis did not avoid urine from male and female deer. Larval and nymphal I. scapularis did not exhibit an arrestment response to urine from male and female white-footed mice, Peromyscus leucopus (Rafinesque), a major host for immature blacklegged ticks. These results suggest that under some conditions, urine from 2 principal host species in certain physiological states does not provide kairomonal cues that aid I. scapularis in contacting these host species.

The cortex, inner and outer medulla, and papilla of kidneys of domestic red deer were analysed. In hydrated animals the urine concentration was found to be 672 +/- 45 mOsm.l-1. The medullary and papillary regions of the kidney were rich in the osmolytes betaine, glycerophosphorylcholine (GPC), inositol and sorbitol, all of which showed a steep rise in concentration from cortex to papilla. The kidney was rich in free amino acids, in particular taurine, glutamate (+glutamine), glycine and alanine, which were present at concentrations sufficient to suggest a possible role as osmolytes.

We examined the effect of progressive nutritional restriction on acid-base balance in seven captive, adult white-tailed deer (Odocoileus virginianus) from 4 February to 5 May 1988 in north central Minnesota (USA). Metabolic acidosis was indicated by low mean blood pH (7.25 to 7.33) in deer throughout the study. Mean urinary pH values declined (P = 0.020) from a mean (+/- SE) baseline of 8.3 +/- 0.1 to 6.7 +/- 0.3 as restriction progressed. Acidemia and aciduria were associated with significant variations in mean blood CO2 (P = 0.006) and pO2 (P = 0.032), serum potassium (P = 0.004) concentrations, and with a significant (P = 0.104) handling date x group interaction in urinary potassium: creatinine values. Mean bicarbonate:carbonic acid ratios were consistently below 20:1 during nutritional restriction. Mean packed cell volume increased (P = 0.019) and serum total protein decreased (P = 0.001); thus there was evidence for progressive dehydration and net protein catabolism, respectively. Blood pCO2, serum sodium, and urinary sodium:creatinine were stable throughout the study. We propose that acidosis and aciduria are metabolic complications associated with nutritional restriction of white-tailed deer.

This study tested the efficacy of assisted reproduction (synchronization of oestrus and intrauterine artificial insemination (AI)) in contributing to the captive propagation of an endangered species, the Eld's deer (Cervus eldi thamin). Semen was collected from males preselected on the basis of under-represented genotype. Motility of spermatozoa after thawing from ejaculates diluted with BF5F extender (8% glycerol), frozen on dry ice in 0.5 ml straws and stored in liquid nitrogen was 60-70%. Intravaginal progesterone-releasing devices (controlled internal drug release, CIDR-type G) were inserted into 20 adult Eld's deer hinds for 14 days. In all hinds, semen (7.5-10 x 10(6) motile spermatozoa per uterine horn) was deposited by laparoscopy performed 70 h after removal of the CIDR device. Ovarian activity, before and after AI, was monitored by analysing pregnanediol-3 alpha-glucuronide (PdG) concentrations in voided urine collected three to seven times per week. During the period of CIDR device insertion, urinary PdG profiles were equal to, or above, normal luteal phase concentrations in all hinds. Within 48 h of device withdrawal, PdG concentrations returned to baseline values in 17 of the 20 females, and the onset of behavioural oestrus occurred at this time in 12 hinds. On the basis of sustained increases in urinary PdG, 9 of the 20 hinds were diagnosed as pregnant by 90 days after AI, all of which delivered offspring after a mean gestation of 241.1 days (range, 235-245). Seven singletons (two females, five males) were born alive and survived, and one singleton and one set of twins were stillborn (three females).(ABSTRACT TRUNCATED AT 250 WORDS)

During winters 1987-1988 (I) and 1988-1989 (II), we assessed the physiological status of moose (Alces alces) residing on the west and east ends of Isle Royale (Michigan, USA) by collecting and chemically analyzing urine deposited in snow (snow-urine) from January to early March. Samples were assayed for urea nitrogen (U), sodium (Na), potassium (K), calcium (Ca), phosphorus (P) and creatinine (C). Throughout both winters, elevated urinary U:C ratios in snow-urine samples collected from east-end moose compared to west-end moose indicated greater dietary energy deprivation and accelerated net catabolism of endogenous protein. Sodium: C ratios were low throughout the study and were similar between moose from both ends of the island, except during the middle of winter I. Greater K:C, P:C and Ca:C ratios in east-end moose compared to west-end moose throughout winter I, and increases in these ratios and U:C in east-end moose from middle to late winter during the second year provided additional evidence of a greater deterioration in condition in east-end moose. The superior nutrition provided to moose on the west end of the island was associated with more developed soils and diverse vegetation and a lower stem density of balsam fir compared to the east end.

We examined the concentration of urinary cortisol and urea nitrogen of five hand-reared mule deer (Odocoileus hemionus) fawns that failed to recover from winter starvation, and compared them to levels found in fawns that recovered. The fawns wintered in fenced pastures stocked with wild deer, and were put back on supplemental feed after losing 15% of their body mass. The five fawns that died began receiving supplemental feed up to 3 wk before death. All continued to lose weight, and were consequently removed from the pasture and fed ad libitum 4 to 10 days before death. In the animals that died, cortisol levels continued to increase regardless of food availability, and were correlated with those of urea nitrogen. Postmortem cortisol and urea nitrogen measurements were significantly greater than concentrations found in the weeks preceding death. We hypothesize that uncontrolled protein catabolism is promoted by high levels of cortisol. These cortisol levels may reach a point at which irreversible multiple-system organ failure occurs, leading to the animal's death.

As a corollary to a more comprehensive study on their ecology, we documented blood and urinary profiles for 10 free-ranging desert mule deer (Odocoileus hemionus crooki) (five males, five females) captured by net-gun shot from a helicopter during February 1988 in Saguaro National Monument, Arizona. Pursuit with the helicopter for netting deer ranged from 3 to 15 min. Blood profiles included seven hematological characteristics and 12 serum chemistries, electrolytes, hormones and enzymes. Urine samples were assayed for urea nitrogen, creatinine, sodium, potassium, calcium and phosphorus. Urinary data were compared as ratios to creatinine. Serum cholesterol was greater (P less than 0.05) in males than females. Pursuit time was correlated with serum non-esterified fatty acids (r = 0.67, P less than 0.05) and influenced urinary specific gravity (r2 = 0.77, P less than 0.004), urea nitrogen: creatinine (r2 = 0.79, P less than 0.005), and potassium: creatinine (r2 = 0.42, P = 0.08) ratios. Increasing specific gravity was related to urinary creatinine concentration (r2 = 0.72, P less than 0.008). All deer exhibited acute adrenal stimulation, accompanied by elevated serum creatine phosphokinase and urinary potassium: creatinine ratios, which were indicative of acute excitement and muscle trauma associated with the capture process. We demonstrated that urinary data are a valuable supplement to serum data in demonstrating effects of intense physical exertion, and both forms of data emphasize the need to assess capture-related excitability as a source of variation in blood and urine characteristics of free-ranging desert mule deer.

The effects of dietary protein, fasting, and refeeding on urinary hydroxyproline of nine captive female white-tailed deer (Odocoileus virginianus) were examined from 23 February to 3 May 1984 in northern Minnesota. In the fasted group, mean hydroxyproline:creatinine (OHP:C) was greater (P less than 0.05) at week 4 compared to baseline at week 0. Between fasted deer and deer fed high protein-high energy (HPHE) and low protein-high energy (LPHE) diets, no difference in OHP:C ratios was detected during the initial 4 wk of the study. Urinary OHP:C ratios were significantly (P less than 0.05) greater in the fasted group during refeeding, concomitant with greater feed consumption and weight gain. There was also a significant (P less than 0.02) time effect in the fasted-refed group; OHP:C ratios increased during these two phases of the study. There was no difference between the HPHE and LPHE fed deer in renal OHP excretion. However, mean OHP:C ratios in yearlings (16.8 +/- 2.2) were greater (P less than 0.001) than in the adults (7.5 +/- 1.2) of those groups, indicating a higher collagen turnover rate. Urinary OHP:C shows potential as an indicator of growth and starvation, and the data presented may serve as reference values.

Three urine sampling techniques were employed in nutritional experiments with captive white-tailed deer (Odocoileus virginianus). Urethral catheterization permitted successful urine collection from females. Furosemide induced urination in male fawns 36.4 +/- 3.1 min (SE) after injection. Significant (P less than 0.05) variation in the responses of individual fawns to this drug were detected. Xylazine hydrochloride induced urination in adult males 91.8 +/- 4.7 min after injection. Significant (P less than 0.01) differences in responses to this drug were detected among individual deer and sample months. The applicability of these urine sampling techniques is discussed.

This study documents glycosuria effects of xylazine and ketamine in eight captive and 19 free-ranging white-tailed deer (Odocoileus virginianus) from January to April 1985. Mean urinary glucose:creatinine ratios in two groups of deer fed high protein-high energy and low protein-low energy diets and in free-ranging deer were 1,000, 719, and 259, respectively. Glucose did not occur in urine of deer immobilized by physical restraint. Glucose:creatinine increased with the time interval between xylazine injection and urine collection in the two groups of captive deer.