Different stressors elicit different responses in the salivary biomarkers cortisol,haptoglobin, and chromogranin A in pigs

Most commonly, salivary cortisol is used in pig stress assessment, alternative salivary biomarkers are scarcely studied. Here, salivary cortisol and two alternative salivary biomarkers, haptoglobin and chromogranin A were measured in a pig stress study. Treatment pigs (n = 24) were exposed to mixing and feed deprivation, in two trials, and compared to untreated controls (n = 24). Haptoglobin differed for feed deprivation vs control. Other differences were only found within treatment. Treatment pigs had higher salivary cortisol concentrations on the mixing day (P < 0.05). Chromogranin A concentrations were increased on the day of refeeding (P < 0.05). Haptoglobin showed a similar pattern to chromogranin A. Overall correlations between the salivary biomarkers were positive. Cortisol and chromogranin A were moderately correlated (r = 0.49, P < 0.0001), correlations between other markers were weaker. The present results indicate that different types of stressors elicited different physiological stress responses in the pigs, and therefore including various salivary biomarkers in stress evaluation seems useful.     

Cortisol release,heart rate,and heart rate variability in transport-naive horses during repeated road transport

Domestic animals are often repeatedly exposed to the same anthropogenic stressors. Based on cortisol secretion and heart rate, it has been demonstrated that transport is stressful for horses, but so far, changes in this stress response with repeated road transport have not been reported. We determined salivary cortisol concentrations, fecal cortisol metabolites, cardiac beat-to-beat (RR) interval, and heart rate variability (HRV) in transport-naive horses (N = 8) transported 4 times over a standardized course of 200 km. Immunoreactive salivary cortisol concentrations always increased in response to transport (P < 0.001), but cortisol release decreased stepwise with each transport (P < 0.05). Concentrations of fecal cortisol metabolites increased from 55.1 ± 4.6 ng/g before the first transport to 161 ± 17 ng/g the morning after (P < 0.001). Subsequent transport did not cause further increases in fecal cortisol metabolites. In response to the first transport, mean RR interval decreased with loading of the horses and further with the onset of transport (1551 ± 23, 1304 ± 166, and 1101 ± 123 msec 1 d before, immediately preceeding, and after 60–90 min of transport, respectively; P < 0.05). Decreases in RR interval during subsequent transports became less pronounced (P < 0.001). Transport was associated with a short rise in the HRV variable standard deviation 2 (P < 0.001 except transport 1), indicating sympathetic activation. No consistent changes were found for other HRV variables. In conclusion, a transport-induced stress response in horses decreased with repeated transport, indicating that animals habituated to the situation, but an increased cortisol secretion remained detectable.     

Monitoring the circadian rhythm of serum and salivary cortisol concentrations in the horse

Daily fluctuations of cortisol concentration in the blood or saliva have been repeatedly reported. However, several contradictions in the existing literature appear on this subject. The present study was performed to definitively establish options for testing adrenocortical function. To the best of our knowledge, this is the first study to evaluate parallel circadian rhythms in salivary and serum cortisol concentrations during a 24-h period. Twenty horses were examined under the same conditions. Blood and saliva samples were taken every 2 h for 24 h to determine the daily changes in cortisol concentrations of saliva and serum at rest and to determine the relationship between salivary and serum cortisol levels. Cosinor analysis of group mean data confirmed a significant circadian component for both serum and salivary cortisol concentrations (P < 0.001 in both cases). The serum cortisol circadian rhythm had an acrophase at 10:50 AM (95% CI, 10:00 AM–11:40 AM), a MESOR of 22.67 ng/mL, and an amplitude of 11.93 ng/mL. The salivary cortisol circadian rhythm had an acrophase at 10:00 AM (95% CI, 9:00 AM–11:00 AM), a MESOR of 0.52 ng/mL, and an amplitude of 0.12 ng/mL. We found a significant but weak association between salivary and serum cortisol concentrations; the Pearson correlation coefficient was 0.32 (P < 0.001). The use of salivary cortisol level as an indicator of hypothalamic-pituitary-adrenal axis activity may be warranted. However, the salivary cortisol levels are more likely to be correlated with free plasma cortisol than with the total plasma cortisol concentration.     

Comparison between blood serum and salivary cortisol concentrations in horses using an adrenocorticotropic hormone challenge

Reasons for performing study: In horses, serum cortisol concentration is considered to provide an indirect measurement of stress. However, it includes both free and bound fractions. The sampling method is also invasive and often stressful. This is not the case for salivary cortisol, which is collected using a more welfare‐friendly method and represents a part of the free cortisol fraction, which is the biologically active form. Objectives: To compare salivary and serum cortisol assays in horses, in a wide range of concentrations, using an adrenocorticotropic hormone (ACTH) stimulation test, in order to validate salivary cortisol for stress assessment in horse. Methods: In 5 horses, blood samples were drawn using an i.v. catheter. Saliva samples were taken using swabs. Cortisol was assayed by radioimmunoassay. All data were treated with a regression method, which pools and analyses data from multiple subjects for linear analysis. Results: Mean ± s.d. cortisol concentrations measured at rest were 188.81 ± 51.46 nmol/l in serum and 1.19 ± 0.54 nmol/l in saliva. They started increasing immediately after ACTH injection and peaks were reached after 96 ± 16.7 min in serum (356.98 ± 55.29 nmol/l) and after 124 ± 8.9 min in saliva (21.79 ± 7.74 nmol/l, P<0.05). Discharge percentages were also different (225% in serum and 2150% in saliva, P<0.05). Correlation between serum and salivary cortisol concentrations showed an adjusted r²= 0.80 (P<0.001). The strong link between serum and salivary cortisol concentrations was also estimated by a regression analysis. Conclusions: The reliability of both RIAs and regression found between serum and salivary cortisol concentrations permits the validation of saliva‐sampling as a noninvasive technique for cortisol level assessment in horses.     

Cortisol release and heart rate variability in sport horses participating in equestrian competitions

Equestrian competitions require both physical activity and mental adaptation in horses. Cortisol, heart rate, and heart rate variability (HRV) are accepted stress parameters and, in this study, have been determined in horses (n = 13) participating in equestrian competitions for up to 3 consecutive days. Participation in competitions caused an increase in salivary cortisol concentrations (e.g., on day 1 from 1.0 ± 0.2 before to 2.2 ± 0.4 ng/mL after the competition, days 1 and 2: P < 0.001, day 3: P < 0.05) and an increase in heart rate (days 1 and 2: P < 0.001, day 3: P = 0.01). A consistent decrease in HRV occurred only in response to the final competition on day 3 (P < 0.01). When horses competing in dressage and show jumping were compared, cortisol release and HRV did not differ between groups, but after the competition, heart rate was lower in dressage than in show jumping horses (P < 0.05). Heart rate increased not only during the actual competition but already when horses were prepared in their stables (e.g., day 1: ?60 minutes, 38.6 ± 2; ?5 minutes, 77 ± 7; competition, 81 ± 10 beats per minute; P < 0.01). In conclusion, participation in equestrian competitions caused an increase in cortisol release and heart rate and a decrease in HRV variables. However, competitions were not a major stressor compared with other anthropogenic challenges such as transport, to which horses are exposed regularly.     

Effect of the change of social environment on the behavior of a captive brown bear (Ursus arctos)

We observed the behavior of a captive sterilized male brown bear before and after the death of his female sibling to investigate the effect of the change of social environment on his behavior and welfare. Observations were carried out by continuous recording during daylight for 6 wk prior to hibernation when the bear was kept with the sibling, and they were repeated 2 years later when the bear was alone (total observation time is 108 h). Feeding, moving, and the total amount of time spent resting (including sleeping and alert inactive) were not affected by the change of social environment. However, when the bear was alone, the percentage of time he spent alert inactive almost trebled (pair, 17.7 ± 3.3%; alone, 48.5 ± 5.5%; P < 0.001), and the time spent sleeping was less than one-third (pair, 51.1 ± 6.1%; alone, 14.2 ± 5.0%; P < 0.001) than when the female was present. The bear spent most of his sleeping time in lateral lying posture (a posture probably associated with rapid eyes movement sleep). The percentage of time dedicated to this posture was significantly reduced after the death of his sibling (pair, 35.9 ± 7.4%; alone, 15.0 ± 5.4%; P < 0.05), whereas the percentage of time spent in quadrupedal posture increased (pair, 17.1 ± 5.4%; alone, 37.8 ± 7.8; P < 0.05). One of the possible reasons for these changes may be an increased risk perception of the bear after the death of his sibling. Our results highlight the importance of social environment and of its changes, which should be carefully considered to maintain captive bears in good welfare conditions.     

Heart rate and salivary cortisol concentrations in foals at birth

Heart rate (HR), HR variability (HRV) and salivary cortisol concentrations were determined in foals (n = 13) during the perinatal phase and until 5 months of age. In the fetus, HR decreased from 77 ± 3 beats/min at 120 min before birth to 60 ± 1 beats/min at 5 min before birth (P <0.01). Within 30 min of birth, HR increased to 160 ± 9 beats/min (P <0.01). Salivary cortisol concentrations immediately after birth were 11.9 ± 3.6 ng/mL and within 2 h increased to a maximum of 52.5 ± 12.3 ng/mL (P <0.01). In conclusion, increases in HR and salivary cortisol concentrations in foals are not induced during parturition, but occur immediately after birth.     

Changes in salivary and plasma cortisol levels in Purebred Arabian horses during race training session

Physical activity and stress both cause an increase in cortisol release ratio. The aim of this study was to evaluate the use of saliva samples for the determination of cortisol concentrations indicating the work‐load level in horses during race training. Twelve Purebred Arabian horses aged 3–5 years were studied during the routine training session. After the warm‐up, the horses galloped on the 800 m sand track at a speed of 12.8 m/s. Three saliva samples, and three blood samples were collected from each horse. Both types of samples were taken at rest, immediately after return from the track and after 30 min restitution. The concentrations of blood lactic acid (LA), and cortisol in saliva and plasma samples were measured and analyzed. Blood LA, plasma and salivary cortisol levels increased significantly after exercise (P < 0.05). Salivary cortisol concentration determined 30 min after the exercise correlated significantly with plasma cortisol level obtained immediately after exercise (P < 0.05) as well as measured 30 min after the end of exercise (P < 0.05). The determination of cortisol concentration in saliva samples taken from racehorses 30 min after the end of exercise can be recommended to use in field conditions to estimate the work‐load in racehorses.     

Plasma hormones, metabolites, milk production, and cholesterol levels in Murrah buffaloes fed with Asparagus racemosus in transition and postpartum period

Ten dry and pregnant Murrah buffaloes were selected to investigate the effect of Asparagus racemosus feeding on hormones, metabolites, milk yield, and plasma cholesterol levels. The treatment groups of buffaloes were fed with A. racemosus (shatavari) @?150?g/day/animal during prepartum and @?300?g/day/animal during the postpartum period. Blood samples collected on ?6, ?4, ?2-week, day of parturition (0), and +2, +4, and +6-week postpartum were analyzed for plasma total cholesterol, triglycerides, HDL, low-density lipoproteins (LDL), prolactin, cortisol, and blood metabolites. Milk samples collected at weekly intervals (+1, +3, +5, and 7?weeks) were analyzed for total milk fat cholesterol. Prepartum plasma cholesterol concentrations were significantly higher in treatment group over the control (P?<?0.05). Mean plasma triglycerides, LDL cholesterol, HDL cholesterol, glucose, and nonesterified fatty acid (NEFA) levels varied nonsignificantly between groups. Plasma prolactin and cortisol concentrations were significantly (P?<?0.01) more in treatment group than in control group. On day of parturition, plasma prolactin, cortisol, LDL, and plasma total cholesterol were higher (P?<?0.01) in treatment group buffaloes in comparison to control group. A. racemosus feeding significantly (P?<?0.01) increased plasma prolactin, cortisol (P?<?0.01), and milk fat cholesterol (P?<?0.05) without affecting total cholesterol, HDL, LDL, glucose, and NEFA concentrations. The buffaloes of treatment group produced more milk (?@?0.526?kg/animal/day) suggesting thereby that A. racemosus is galactopoietic. It was concluded that feeding of A. racemosus increases plasma prolactin and cortisol and decreased plasma total cholesterol and LDL concentration.     

Feed efficiency and body composition are related to cortisol response to adrenocorticotropin hormone and insulin-induced hypoglycemia in rams

Metabolic rate and energy consumption increase through the activation of the hypothalamic-pituitary-adrenal axis when an animal is exposed to a stressor. Residual feed intake (RFI) as a measure of efficiency has been shown to be related to exogenous adrenocorticotropin hormone (ACTH)-stimulated cortisol concentrations, which is indicative of the relationship between an animal's response to stress and the efficiency with which the energy is used for growth and production. In this study, we tested the hypothesis that sheep with low post-ACTH serum cortisol concentration relative to the other sheep in the flock have lower RFI values and lower cortisol concentrations following insulin-induced hypoglycemia. Adrenocorticotropin hormone (2.0 μg/kg body weight)-stimulated cortisol concentrations were measured in 100 sheep. The extreme responders were selected (n = 12 high cortisol, n = 12 low cortisol), and feed efficiency and body composition parameters were measured. A second ACTH challenge and an insulin challenge were administered. More efficient sheep (more negative RFI value) were found to have lower (P < 0.05) cortisol concentrations following both an ACTH challenge and an insulin challenge. Low-cortisol sheep (low response to ACTH or insulin) were found to have a lower (P < 0.05) proportion of fat tissue in comparison to the high-cortisol animals. These data clearly indicate that an animal's response to exogenous ACTH or insulin-induced hypoglycemia as a stressor is related (P < 0.05) to efficiency of energy use when measured as RFI. These data have important implications in enabling identification of animals that are superior in terms of feed efficiency and for understanding the physiological mechanisms underlying efficiency of energy use.     

Changes in cortisol release and heart rate variability in sport horses during long-distance road transport

It is widely accepted that transport is stressful for horses, but only a few studies are available involving horses that are transported regularly and are accustomed to transport. We determined salivary cortisol immunoreactivity (IR), fecal cortisol metabolites, beat-to-beat (RR) interval, and heart rate variability (HRV) in transport-experienced horses (N = 7) in response to a 2-d outbound road transport over 1370 km and 2-d return transport 8 d later. Salivary cortisol IR was low until 60 min before transport but had increased (P < 0.05) 30 min before loading. Transport caused a further marked increase (P < 0.001), but the response tended to decrease with each day of transport. Concentrations of fecal cortisol metabolites increased on the second day of both outbound and return transports and reached a maximum the following day (P < 0.001). During the first 90 min on Day 1 of outbound transport, mean RR interval decreased (P < 0.001). Standard deviations of RR interval (SDRR) decreased transiently (P < 0.01). The root mean square of successive RR differences (RMSSD) decreased at the beginning of the outbound and return transports (P < 0.01), reflecting reduced parasympathetic tone. On the first day of both outbound and return transports, a transient rise in geometric HRV variable standard deviation 2 (SD2) occurred (P < 0.01), indicating increased sympathetic activity. In conclusion, transport of experienced horses leads to increased cortisol release and changes in heart rate and HRV, which is indicative of stress. The degree of these changes tended to be most pronounced on the first day of both outbound and return transport.     

The impact of dystocia on dairy calf health,welfare, performance and survival

Up to one-third of dairy calves are born after dystocia and this is a major cause of calf mortality. This study investigated the neonatal physiology, survival, health and subsequent growth of dairy calves following dystocia and is the first longitudinal study to analyse multiple effects and to look beyond the perinatal period.A total of 455 live born Holstein calves (N: No assistance, n = 360; FN: Farmer assistance but normally presented calf, n = 82; FM: Farmer assistance of malpresented calf, n = 13) were followed from birth to first service (heifers) or until leaving the farm (bulls). Compared to N calves, FN and FM animals had higher salivary cortisol concentrations at day 1 (P < 0.001) and FN calves had lower passive immune transfer (P = 0.03). Dystocia had no biologically significant impact on rectal temperature throughout the first 4 days (P > 0.05). During the first 60 days, FM calves had a higher proportion of days with non-routine health treatments (P < 0.05) and, by the time of weaning, mortality in FN and FM heifers was higher than in N calves (2.8×; P < 0.01). However, in surviving calves, growth to first service was not affected by dystocia category (P > 0.05).Calves which survive dystocia experience lower passive immunity transfer, higher mortality and higher indicators of physiological stress. Such calves have poorer welfare in the neonatal period and possibly beyond. Strategies need to be implemented to improve the subsequent health and welfare of such calves and to lower the incidence of dystocia.     

Temporal pattern and effect of sex on lipopolysaccharide-induced stress hormone and cytokine response in pigs

The temporal pattern and sex effect of immune and stress hormone responses to a lipopolysaccharide (LPS) challenge were assessed using a pig model. Secretion of the pro-inflammatory cytokines tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 increased in a time-dependent manner following LPS infusion. There was also a time-dependent increase in secretion of the stress-related hormones cortisol, epinephrine (E), and norepinephrine (NE) following LPS, with peak concentrations attained within 30 min. The magnitude of the TNF-α and IL-1β responses were both positively associated (P < 0.05) with the magnitude of cortisol response following LPS, whereas serum IL-1β and IL-6 were positively correlated with the magnitude of E and NE responses following LPS. Acute-phase protein production was also time-dependently increased following LPS. The concentration of immune cells in circulation was decreased (P < 0.05) at 5.5 h post-LPS and negatively correlated with pro-inflammatory cytokine production. By 24 h post-LPS, immune cell counts increased (P < 0.05) and were positively associated with both pro-inflammatory cytokine and stress hormone production. The amplitude of pro-inflammatory cytokine response following LPS was affected (P < 0.05) by sex classification; however, the magnitude of elevated cytokine concentrations was not. The magnitude of the NE response, but not of the E and cortisol responses, to LPS was influenced by sex (P < 0.05). Similar to the pro-inflammatory cytokines, the magnitude of exposure to the stress hormones following LPS was not influenced by sex. The production of serum amyloid A (SAA) was influenced by sex, with barrows producing more SAA than gilts at 24 h post-LPS (P < 0.05). Collectively, these results demonstrate sex-specific, concomitant temporal changes in innate immune- and stress-related hormones.     

Environmental enrichment and social rank affects the fear and stress response to regular handling of dairy goats

Human–animal interactions can have result in fear and stress for the animals and can affect negatively their welfare and productivity. Environmental enrichment techniques can be a tool to reduce this fear response to handlers. The aim of this study was to analyze the effects of environmental enrichment and social rank on fear and stress response to handling of dairy goats. Thirty Saanen dairy goats (3.5-5 years of age) were observed 6 hours daily for 16 consecutive days. Behavior sampling was used to record all events of agonistic interactions, and index of success was calculated for each goat. Two groups, control (CO) and enriched (EN), of 12 goats each (6 high rank [HR] and 6 low rank [LR]), were then observed during a 4-phase handling test. Reactivity behaviors were recorded using focal sampling, and plasma cortisol was assessed in all goats. An analysis of variance (ANOVA) was used to assess the effect of treatment (EN and CO) and social rank (HR and LR) on behavior, and a multivariate ANOVA for repeated measures was used to assess the effect of treatment and social rank on cortisol levels in response to handling. The EN group and HR goats had a longer distance to the handler (P < 0.01 and P < 0.05, respectively); however, it took longer for the goats in the CO group to be caught (P < 0.01). LR goats in the EN group showed higher levels of aggression to the handler than HR goats in the EN and CO groups as well as LR goats in the CO group (P < 0.01). On average, EN goats had higher cortisol values than in the CO (P < 0.05), and the HR goats had significantly higher cortisol values than the LR animals (P < 0.05). In general, goats in the EN group had a more excited reaction than the CO group that could be related to a cognitive state derived from the effect of the enrichment. These results have important implications for animal husbandry and help to broaden the literature on the effects of environmental enrichment and dominance rank on goat handling.     

Evaluation of rumen-protected choline additive to diet on productive performance of male Zaraibi growing goats during hot summer season in Egypt

Twenty Zaraibi goat bucks were used in this experiment which lasted 3 months during summer season of Egypt. The animals were divided randomly into two equal groups. The first group was kept without treatment as control while in the second group, rumen-protected choline (RPC) at the level of 20 g/buck/day was added to the concentrate feed mixture at the morning feeding. RPC additives to diet of Zaraibi goat bucks during the period of hot summer season increased (P < 0.01) total gain and average daily gain compared to the control group. RPC increased (P < 0.05) dry matter intake and feed conversion while water intake was not affected by RPC additives. RPC increased (P < 0.05) red and white blood cell (RBC × 10⁶, WBC × 10³) counts and hemoglobin concentration and hematocrit percentage. RPC increased total protein (P < 0.05), globulin, and γ-globulin (P < 0.01). On the other hand, total lipids, total cholesterol, and triglyceride concentrations decreased (P < 0.05 and P < 0.05) while phospholipids, glucose, and choline concentrations increased (P < 0.01) due to RPC supplementation. RPC increased (P < 0.01) thyroxin and triiodothyronine, increased (P < 0.05) testosterone levels, and decreased (P < 0.01) cortisol level compared with control bucks. It is concluded that dietary RPC at the rate of 20 g daily is required for growing male goats, especially, under heat stress conditions of summer season in Egypt and showed the best results concerning the growth, feed conversion, blood metabolites, and economic efficiency.     

Salivary Cortisol Concentrations in Healthy Dogs and Dogs with Hypercortisolism

Background: Measurement of salivary cortisol is a useful diagnostic test for hypercortisolism (HC) in humans. Objectives: To determine whether measurement of salivary cortisol concentration is a practical alternative to plasma cortisol to diagnose HC, to validate the use of salivary cortisol, and to examine the effect of time of day and sampling location on salivary cortisol. Animals: Thirty healthy dogs and 6 dogs with HC. Methods: Prospective, observational clinical trial including healthy volunteer dogs and dogs newly diagnosed with HC. Salivary and plasma cortisol concentrations were measured with an immunoassay analyzer. Intra‐ and interassay variability, linearity, and correlation between salivary and plasma cortisol concentrations were determined. Results: The required 300 μL of saliva could not be obtained in 88/326 samples from healthy dogs and in 15/30 samples from dogs with HC. The intra‐assay variability for measurement of salivary cortisol was 5–17.7%, the interassay variability 8.5 and 17.3%, and the observed to expected ratio 89–125%. The correlation (r) between salivary and plasma cortisol was 0.98. The time of day and location of collection did not affect salivary cortisol concentrations. Dogs with HC had significantly higher salivary cortisol values than healthy dogs (10.2 ± 7.3 nmol/L versus 1.54 ± 0.97 nmol/L; P < .001). Conclusions and Clinical Importance: The ROCHE Elecsys immunoassay analyzer correctly measured salivary cortisol in dogs. However, a broad clinical application of the method seems limited, because of the large sample volume required.     

Haematological findings in captive dolphins and whales

Objective To examine haematological features in five species of healthy, captive marine mammals.
Animals Twenty bottlenose dolphins ( Tursips truncatus ), seven Pacific white-sided dolphins ( Lagenorhynchus obliquidens ), five Risso dolphins ( Grampus griseus ) and five false killer whales ( Pseudorca crassidens ).
Results and conclusion The red blood cell count was 4.21 × 10¹²/L in bottlenose dolphins, 5.32 × 10¹²/L in Pacific white-sided dolphins, 4.35 × 10¹²/L in Risso dolphins and 4.43 × 10¹²/L in false killer whales. The haemoglobin concentration was 1.51 g/L and packed cell volume 44.7% in bottlenose dolphins; the corresponding values were 1.71 g/L and 48.9% in Pacific white-sided dolphins, 1.72 g/L and 49.4% in Risso dolphins, and 1.52 g/L and 47.8% in false killer whales. The white blood cell count was 7.097 × 10⁹/L in bottlenose dolphins, 5.928 × 10⁹/L in Pacific white-sided dolphins, 5.001 × 10⁹/L in Risso dolphins and 7.921 × 10⁹/L in false killer whales. There were no significant differences in these values among bottlenose dolphins and Pacific white-sided dolphins. The proportion of eosinophils in the differential leukocyte count ranged from 10.3% to 11.5% in bottlenose dolphins, Pacific white-sided dolphins and false killer whales, but was only 0.4% in Risso dolphins. The eosinophilic granules were larger in Risso dolphins and false killer whales than in bottlenose and Pacific white-sided dolphins.     

Bartonella species detection in captive, stranded and free-ranging cetaceans

We present prevalence of Bartonella spp. for multiple cohorts of wild and captive cetaceans. One hundred and six cetaceans including 86 bottlenose dolphins (71 free-ranging, 14 captive in a facility with a dolphin experiencing debility of unknown origin, 1 stranded), 11 striped dolphins, 4 harbor porpoises, 3 Risso's dolphins, 1 dwarf sperm whale and 1 pygmy sperm whale (all stranded) were sampled. Whole blood (n = 95 live animals) and tissues (n = 15 freshly dead animals) were screened by PCR (n = 106 animals), PCR of enrichment cultures (n = 50 animals), and subcultures (n = 50 animals). Bartonella spp. were detected from 17 cetaceans, including 12 by direct extraction PCR of blood or tissues, 6 by PCR of enrichment cultures, and 4 by subculture isolation. Bartonella spp. were more commonly detected from the captive (6/14, 43%) than from free-ranging (2/71, 2.8%) bottlenose dolphins, and were commonly detected from the stranded animals (9/21, 43%; 3/11 striped dolphins, 3/4 harbor porpoises, 2/3 Risso's dolphins, 1/1 pygmy sperm whale, 0/1 dwarf sperm whale, 0/1 bottlenose dolphin). Sequencing identified a Bartonella spp. most similar to B. henselae San Antonio 2 in eight cases (4 bottlenose dolphins, 2 striped dolphins, 2 harbor porpoises), B. henselae Houston 1 in three cases (2 Risso's dolphins, 1 harbor porpoise), and untyped in six cases (4 bottlenose dolphins, 1 striped dolphin, 1 pygmy sperm whale). Although disease causation has not been established, Bartonella species were detected more commonly from cetaceans that were overtly debilitated or were cohabiting in captivity with a debilitated animal than from free-ranging animals. The detection of Bartonella spp. from cetaceans may be of pathophysiological concern.     

Sexual maturity,seasonal estrus,and gestation in female Indo-Pacific bottlenose dolphins Tursiops aduncus inferred from serum reproductive hormones

Reproductive hormones in serum concentrations of progesterone, estradiol, and testosterone in female Indo-Pacific bottlenose dolphins (Tursiops aduncus, n = 12) housed in Ocean Park Hong Kong were investigated in the present study. Results showed that, onset of puberty of captive Indo-Pacific bottlenose dolphins was at 5 years while sexual maturity was at 6. Average serum progesterone concentrations in non-pregnant sexually mature individuals was 0.33 (0.25–0.97) ng/mL (interquartile), significantly higher than in immature ones 0.26 (0.25–0.38) ng/mL. This study found significant difference in serum estradiol concentrations between individuals at the onset of puberty (9.5 ± 1.7 pg/mL, ±SD) and not (below detection limit 9 pg/mL). A slightly seasonal breeding pattern, with progesterone values tend to be higher from February to October (0.38 [0.25–1.07] ng/mL) was inferred. During pregnancy, serum progesterone concentrations range from 10.54 ± 8.74 ng/mL (indexed month post-conception [IMPC] 0) to 25.49 ± 12.06 ng/mL (IMPC 2), and display a bimodal pattern with 2 peaks in early- (25.49 ± 12.06 ng/mL, IMPC 2) and late-pregnancy (21.71 ± 10.25 ng/mL, IMPC 12), respectively. Serum estradiol concentrations can seldom be detected in early-pregnancy and increase constantly in mid- (9.45 ± 1.83 pg/mL) and late-pregnancy (11.88 ± 3.81 pg/mL), with a spike (15.45 ± 6.78 pg/mL) 1 month prior to delivery. Serum testosterone concentrations elevate significantly in IMPC 7 (0.36 ± 0.10 ng/mL) compared to other months (0.16 ± 0.10 ng/mL) of the year. The present study provides normal concentration profiles for some reproductive hormones in female Indo-Pacific bottlenose dolphins and can contribute to the breeding monitoring of this species. Also, our study would shed further light on the reproductive physiology of small cetaceans.     

The Potential Role of Training Sessions on the Temporal and Spatial Physiological Patterns in Young Friesian Horses

This study compares the circulating adrenocorticotrophin hormone (ACTH), cortisol, lactate, glucose, heart rate, respiratory rate, rectal temperature, and blood count values in initially 2-year-old horses subjected to dressage training schedule during three consecutive days per 2 weeks. Sixteen healthy Friesian horses were used and were considered dressage group. Six healthy young horses not involved in training programs were used as control group. Blood sampling were collected from the jugular vein in baseline condition (dressage group and control group) and after exercise, within 5 minutes of the end of the training session (dressage group). Compared to baseline values, results showed higher ACTH concentrations after the first day of the first training week (P < .005) and after the third day of the second week (P < .005); higher lactate concentrations after the second and the third day of the second week (P < .01); lower glucose concentrations after the third day of the first week (P < .01); higher HR, RR, and RT values and lower PLT count after different time points during both training weeks. One-way ANOVA showed significant training effect for ACTH (F = 7.605; P < .0001) and glucose (F = 3.505; P < .001) concentrations over time points. Two-way ANOVA showed a significant effect of dressage training sessions between the first and the second week for ACTH (F = 6.508; P < .001) and cortisol (F = 5.559; P < .0001) concentrations. From obtained data, it seems that the use of ACTH and cortisol changes for the assessment of effects of training in initially 2-year-old horses could be an ideal measure of quantitative and qualitative stress responses. The quantification at the same time of functional responses to stressful stimuli may offer a more objective measurement of dressage training effects.