Neville Pitts

Seasonal patterns of hormones, macroparasites, and microparasites in wild African ungulates: the interplay among stress, reproduction, and disease.

Sex hormones, reproductive status, and pathogen load all affect stress. Together with stress, these factors can modulate the immune system and affect disease incidence. Thus, it is important to concurrently measure these factors, along with their seasonal fluctuations, to better understand their complex interactions. Using steroid hormone metabolites from fecal samples, we examined seasonal correlations among zebra and springbok stress, reproduction, gastrointestinal (GI) parasite infections, and anthrax infection signatures in zebra and springbok in Etosha National Park (ENP), Namibia, and found strong seasonal effects. Infection intensities of all three GI macroparasites examined (strongyle helminths, Strongyloides helminths, and Eimeria coccidia) were highest in the wet season, concurrent with the timing of anthrax outbreaks. Parasites also declined with increased acquired immune responses. We found hormonal evidence that both mares and ewes are overwhelmingly seasonal breeders in ENP, and that reproductive hormones are correlated with immunosuppression and higher susceptibility to GI parasite infections. Stress hormones largely peak in the dry season, particularly in zebra, when parasite infection intensities are lowest, and are most strongly correlated with host mid-gestation rather than with parasite infection intensity. Given the evidence that GI parasites can cause host pathology, immunomodulation, and immunosuppression, their persistence in ENP hosts without inducing chronic stress responses supports the hypothesis that hosts are tolerant of their parasites. Such tolerance would help to explain the ubiquity of these organisms in ENP herbivores, even in the face of their potential immunomodulatory trade-offs with anti-anthrax immunity.

Effects of environmental endocrine disruptors, including insecticides used for malaria vector control on reproductive parameters of male rats.

The male reproductive system is sensitive to endocrine disrupting chemicals (EDCs) during critical developmental windows. Male Sprague-Dawley rats were exposed in utero-, during lactation- and directly to 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (DDT), 1,1,-dichloro-2,2-bis(p-chlorophenyl)ethylene (DDE) and a mixture of DDT, deltamethrin (DM), p-nonylphenol (p-NP) and phytoestrogens, at concentrations found in a malaria-area. After dosing for 104 days, histological assessments and reproductive-endpoints were assessed. The anogenital distance (AGD) (P=0.005) was shorter in the mixture-exposed group, while the prostate mass (P=0.018) was higher in the DDT-exposed group. A higher testicular mass and abnormal histology was observed in the DDT-(P=0.019), DDE-(P=0.047) and mixture-exposed (P<0.005) groups. This study shows that in utero-, lactational- and direct exposure to EDCs present in a malaria-area negatively affects male reproductive parameters in rats. These findings raise concerns to EDC-exposures to mothers living in malaria-areas and the reproductive health of their male offspring.

Pharmacokinetics and effects of succinylcholine in African elephant (Loxodonta africana) and impala (Aepyceros melampus)

The phenomenon of slow onset of succinylcholine (Sch) effect in elephants was investigated by analyzing blood concentrations of Sch and its metabolite choline in elephant and impala. To assess whether the slow onset phenomenon is related to the pharmacokinetics of Sch following i.m. administration, we analyzed the time course of plasma concentrations of intact drug and its metabolite and determined its pharmacological effects. Blood samples were obtained from anaesthetized elephant (n=6) and impala (n=7) following i.m. administration of a lethal dose of Sch. Time from Sch injection to onset of apnoea and to death was significantly longer for elephant than impala (mean+/-S.D. apnoea 4.4+/-1.5 and 2.3+/-0.9 min, respectively; death 32.6+/-7.3 and 6.2+/-3.4 min, respectively). The C(max) was not different between elephants and impala (20.3+/-7.9 vs. 14.4+/-6.8 nmol ml-1, respectively) but the t(max) was significantly longer for elephants (23.0+/-7.6 vs. 3.7+/-2.2 min). Analysis of the plasma Sch and choline concentrations over time revealed that the relative amount of Sch entering the circulation within the first 30 s after i.m. injection is greater for impala than elephant. No greater rate in the plasma hydrolysis of Sch in elephant compared to impala was apparent.

Validating a human biotelemetry system for use in captive blue wildebeest (Connochaetes taurinus)

We fitted two blue wildebeest (Connochaetes taurinus) with modified versions of the Equivital™ EQ02 wireless monitoring system to evaluate if the device could accurately measure heart rate and respiration rate in this species whilst anaesthetized as well as whilst fully conscious in captivity. Whilst under anaesthesia, we monitored each animals heart rate and respiration rate using the Equivital™ biotelemetry belt, a Cardell(®) veterinary monitor and manual measurements. The animals were also administered doxapram hydrochloride (Dopram(®) ) and adrenaline intravenously at different times to stimulate changes in respiration and heart rate, respectively. Once 30 minutes of monitoring was completed, we reversed the anaesthetic and left the animals in captivity for 24 hours whilst wearing the Equivital™ belts. After 24 hr, we repeated the anaesthesia and monitoring as well as the administration of the doxapram hydrochloride and adrenaline. Intraclass Correlation Coefficients (ICC) calculated between all three monitoring methods showed moderate to excellent agreements for heart rate on both days (ICC: 0.73-0.98). ICCs calculated between the three methods for respiration rate showed good to excellent agreement between the Equivital belt and the other two methods (0.82-0.92) with the exception of occasions when only poor to fair agreements were found between the Cardell(®) measurements and manual measurements. Heart rate and respiration rate were also found to increase with motion while animals were in captivity. The results indicate that a modified version of the Equivital™ EQ02 system can be used as a potential biotelemetry device for measuring heart and respiration rate in captive blue wildebeest.

Disparate plasma cortisol concentrations in sexually abused female children from Johannesburg, South Africa

A growing body of research indicates that a bidirectional response to a stressor may occur in maltreated children and may be associated with later life psychopathology. However, few studies have investigated stress reactivity in children when they first present to a sexual abuse clinic. Thus, in order to evaluate whether HPA axis dysregulation would be evident at first presentation to a sexual abuse clinic in young girls (n = 26), between the ages of 6-12 years old, blood samples were obtained immediately following examination at a forensic sexual abuse clinic and from the matched control group of children (n = 14; 10.1 ± 0.8) immediately following a bone density scan. Stratification of the sexually abused group into those children who were reportedly abused by a stranger and had no other family stressors (n = 15, 10.4 ± 1.8) and those children whose parents reported abuse of the child by a stranger and other family stressors (n = 11; 9.5 ± 1.8) revealed differences in stress reactivity. Plasma concentrations, of the children from the forensic clinic, were significantly increased in children who reported abuse by a stranger only (322.3 ± 117.4 nmol/l) and significantly decreased in children whose histories indicated sexual abuse by a stranger and other family stressors (149.6 ± 39.7 nmol/l) when compared to the control group (225.5 ± 47.5 nmol/l). In conclusion, following sexual abuse and a secondary stressor, the forensic examination, there is evidence of divergent cortisol responses in the stratified clinical group of children.

Noninvasive measures of stress response in African buffalo (Syncerus caffer) reveal an age-dependent stress response to immobilization

Fecal glucocorticoid metabolites (FGMs) are commonly used as indicators of an animals stress response in behavioral and ecophysiological studies. FGM assays provide a noninvasive and efficient means of assessing adrenocortical activity. We used 12 African buffalo (Syncerus caffer) temporarily maintained in an enclosure to evaluate 2 commercially available FGM assays as tools for assessing stress levels in buffalo in field studies. We also used the experiment to assess potential adverse effects of immobilizations on the study animals. Buffalo responded rapidly to stimulation with adrenocorticotropic hormone (ACTH), which stimulates the adrenal cortex to secrete glucocorticoids. ACTH-stimulated buffalo had higher plasma cortisol concentrations than saline-injected controls between 5 min and 1 h after injection. The ACTH-induced plasma cortisol peak was detectable in FGMs at 10–20 h post-injection. Both of the commercial test kits we evaluated were capable of detecting the ACTH-induced peak in FGM. However, the radioimmunoassay delivered more consistent detection across weeks than the enzyme immunoassay. We tested whether immobilization and handling elicited a stress response detectable by FGM, by comparing immobilized, saline-injected buffalo with controls that were not immobilized or handled. Adult buffalo mounted a stress response to immobilization and handling, whereas subadults did not, suggesting an age-related difference in response to chemical immobilization. Our study validates use of commercially available kits for quantifying FGMs under field conditions.

THE EFFECT OF A SLOW-RELEASE FORMULATION OF ZUCLOPENTHIXOL ACETATE (ACUNIL®) ON CAPTIVE BLUE WILDEBEEST (CONNOCHAETES TAURINUS) BEHAVIOR AND PHYSIOLOGICAL RESPONSE

Abstract The study investigated the effect of a slow-release formulation of zuclopenthixol acetate (Acunil®) on blue wildebeest (Connochaetes taurinus) in captivity. Two groups of trials were conducted using either Acunil or a placebo (control). Animals (Acunil: n = 17; placebo: n = 12) were observed for a 12-hr period before the administration of Acunil or the placebo (pretreatment). After 24 hr, animals were administered Acunil (1.5 mg/kg) or a placebo (1.0–3.0 ml of sterile water) and observed again for 12 hr (posttreatment). During both treatments, animals were stimulated every 2 hr for 1 min by a person entering the enclosure (referred to as periods of stimulation). Behavioral observations and continuous heart rate, respiration rate, and motion measurements were taken throughout. Animals treated with Acunil spent more time lying with their heads folded back, eating and standing with their heads down, and less time being vigilant and exploring while walking around. Animals treated with the placebo also spent less time being vigilant and more time lying with heads up. Animals treated with Acunil groomed less while standing and performed less head shaking; no such changes were observed in the control group. Neither Acunil nor the placebo had any effect (P > 0.05) on heart rate. However, overall mean respiration rate was lowered (P = 0.02) when animals were treated with Acunil (pretreatment: 14.5 ± 0.82 breaths/min; posttreatment: 12.5 ± 0.83 breaths/min). Acunil also caused a lowered (P < 0.05) respiration rate during periods when animals were stimulated (pretreatment: 16.2 ± 0.87 breaths/min; posttreatment: 13.7 ± 0.87 breaths/min) and when animals were trotting and being vigilant. No such changes were observed with the placebo. Both placebo- and Acunil-treated animals spent more time being stationary during periods of stimulation. However, Acunil-treated animals also spent less time moving fast when they were stimulated.

83. Disrupted attachment: Allostatic load one day post weaning

IL-12 expression was suppressed for non-inhibited, but not for inhibited, monkeys. These results suggest that the modulating effect of glucocorticoids on cytokine gene expression is tissue-specific (APCs vs. T-cells). Evidence is emerging that IL-12 can influence function of regulatory T-cells (TReg), and the failure of glucocorticoids to suppress IL-12, but not IFN-c expression, in inhibited animals suggests that the influence of behavioral inhibition on atopic disorders may be related to impairment in the regulatory function in the TReg population.

In vitro succinylcholine hydrolysis in plasma of the African elephant (Loxodonta africana) and impala (Aepyceros melampus)

In elephants the time lapsed from i.m. injection of an overdose of the muscle relaxant succinylcholine (SuCh) until death, is significantly longer than in impala. To determine a difference in the rate of SuCh hydrolysis, once the drug enters the circulation, contributes to this phenomenon we have measured the rate of hydrolysis of SuCh in elephant and impala plasma, and by elephant erythrocytes. Rate of hydrolysis was determined by incubating SuCh in plasma or erythrocyte lysate at 37 degrees C and quantifying the choline produced. Plasma SuCh hydrolytic activity in elephant plasma (12.1+/-1.7 Ul(-1) mean+/-S.D.; n=9) was significantly higher than it was in impala plasma (6.6+/-0.6 Ul(-1); n=5), but were approximately 12 and 21 times lower, respectively, than in human plasma. Elephant erythrocyte lysate had no SuCh hydrolytic activity. Applying this data to previous studies, we can show that the ratio of SuCh absorption to SuCh hydrolysis is expected to be 1.25:1 and 1.41:1 for elephants and impala respectively. It will thus take at least 1.7 times longer for elephant to achieve a plasma SuCh concentration similar to that in impala. We conclude that a more rapid hydrolysis of SuCh in elephant plasma is one factor that contributes to the longer time to death compared to impala.