The influence of physiological status on the reproductive behaviour of humpback whales (Megaptera novaeangliae).

For most cetacean species, there is little known about how an individual's physiology influences its behaviour. Humpback whales (Megaptera novaeangliae) are a good candidate to examine such links as they have a well-described distribution and behaviour, can be consistently sampled using remote biopsy systems, and have been the subject of several previous endocrine studies. The objective here was to examine whether a female humpback whale's social state (i.e. escorted by a male or not) is related to her endocrine condition, and whether male dominance ranking is related to testosterone levels. Skin and blubber biopsies were collected from the east and west Australian humpback whale populations in 2010-2016 (n = 252) at multiple times throughout the winter-spring breeding season. Steroid hormones were extracted from blubber and concentrations of progesterone (a marker for pregnancy), testosterone (a marker of male testicular activity) and oestradiol (a potential marker of ovarian activity) measured using enzyme-immunoassays. Principal escorts-the dominant males in mixed sex groups-had significantly higher blubber testosterone levels (mean ±â€¯SE; 1.43 ±â€¯0.20 ng/g wet weight) than subordinate, secondary escorts (0.69 ±â€¯0.06 ng/g wet weight). Females that were escorted by males typically possessed elevated blubber oestradiol levels (1.96 ±â€¯0.25 ng/g wet weight; p = 0.014); few were considered to be pregnant (p = 0.083). 'Unescorted' females displayed characteristically lower blubber oestradiol levels (0.56 ±â€¯0.06 ng/g wet weight). Together, these results are consistent with 'challenge hypothesis' theory and suggest the existence of associated reproductive patterns in humpback whales.

Evaluation of respiratory vapour and blubber samples for use in endocrine assessments of bottlenose dolphins (Tursiops spp.).

Blubber and respiratory vapour ('blow') are now commonly used for endocrine studies on cetaceans, primarily because they can be obtained using minimally invasive methods. For many species, these samples have yet to be validated for these purposes. The objective of this study was to examine the performance of blow and blubber hormone monitoring, relative to serum hormone monitoring, for evaluating the reproductive and adrenal condition of captive bottlenose dolphins (Tursiops spp.). Eighteen bottlenose dolphins were sampled five times for serum and blow and twice for blubber throughout a one-year period. Concentrations of progesterone, testosterone, oestradiol and cortisol were measured in each sample type. Hormone levels were examined in relation to dolphin age, sex, reproductive status, season, time of sample collection (morning/afternoon) and collection type (in- or out-of-water sampling). Patterns in hormone levels were similar for serum and blubber. For instance, in both sample types, progesterone levels were significantly higher in pregnant (serum: 34.10 ±â€¯8.64 ng/mL; blubber: 13.01 ±â€¯0.72 ng/g) than in non-pregnant females (serum: 0.32 ±â€¯0.09 ng/mL; blubber: 1.17 ±â€¯0.10 ng/g). This pattern was not detected in blow, primarily because seawater contamination, nylon sampling materials and variable sample volumes influenced measured concentrations. In addition, the respiratory water content of a blow sample is known to affect measured hormone levels. Two methods were trialled to control for variability in sample volumes and dilution: (1) normalising blow hormone concentrations relative to urea nitrogen levels (a potential endogenous standard), and (2) measuring the relative proportions (i.e. ratios) of blow hormones. These correction measures had little influence on blow hormone results. Further refinement of blow hormone monitoring methods is required before they can be used for reproductive or adrenal assessments of bottlenose dolphins. Blubber, on the other hand, should be a suitable proxy for serum when attempting to classify pregnancy status and male maturity in these species.

Behavioral and endocrine responses to season and social dynamics of captive male southern hairy-nosed wombats (Lasiorhinus latifrons).

Although southern hairy-nosed wombats (SHN wombats; Lasiorhinus latifrons) rarely breed in captivity, further knowledge of their reproductive physiology and behavior is likely to improve their breeding potential. This study examined the effect of seasonal variation and changes in social dynamics on the physiology and behavior of a captive population of male SHN wombats (n = 6). Seasonal changes in urinary testosterone metabolites (UTM), urinary cortisol metabolites (UCM), qualitative estimates of spermatorrhoea (QS), aggressive behavior and reproductive behavior were measured over an 11-month period. While there was no effect of month on QS (GLM ANOVA, P = 0.27), reproductive behavior (GLM ANOVA, P = 0.19) or aggressive behavior (Tukey pairwise comparisons), the secretion of UTM (GLM ANOVA, P = 0.051) was only marginally affected by season, compared to that reported for wild male SHN wombats. Mean UCM concentrations of July and August 2016 were significantly higher than those between October 2015 and January 2016 (Tukey pairwise comparisons). To examine social dynamics, two trials of animal positioning exchange with the enclosure system were implemented and behavioral data were examined for each trial over a six week period; UTM, UCM and general behaviors (n = 27) were measured for each trial. Neither UTM nor UCM concentration varied significantly (P ≥ 0.45) before and after the exchanges. "Scratching" decreased at the group level following the animal exchange in both trials, suggesting reduction in self-grooming may be a behavioral response to novel stimuli. UCM and UTM concentrations were both positively correlated with "standing still" and "body rub" behaviors. This may be evidence of a hormonal control of a "freezing behavioral response" to external stimuli and marking behavior, respectively. As there was no evidence that changing the social dynamics affected reproductive or agonistic behavior or hormone concentrations, it was concluded that captive male wombats in this study showed reduced reproductive seasonality compared to wild wombats and that animal exchange resulted in a behavioral response to novel stimuli but was not sufficient to affect testosterone or cortisol secretion, within the context of our study.

Cryopreservation of saltwater crocodile (Crocodylus porosus) spermatozoa.

The aim of the present study was to develop a protocol for the successful cryopreservation of Saltwater crocodile spermatozoa. Sperm cells were frozen above liquid nitrogen vapour in phosphate-buffered saline (PBS) containing either 0.3M trehalose, 0.3M raffinose or 0.3M sucrose and compared with glycerol (0.3-2.7M). Although the highest levels of mean post-thaw motility were observed following cryopreservation in 0.3M trehalose (7.6%) and 0.3M sucrose (7.3%), plasma membrane integrity (PI) was best following cryopreservation in 2.7M glycerol (52.5%). A pilot study then assessed the cytotoxicity of glycerol and sucrose prior to cryopreservation and revealed no loss of survival when spermatozoa were diluted in 0.68M glycerol or 0.2-0.3M sucrose once cryoprotectants were washed out with PBS or Biggers, Whitten and Whittingham medium containing sperm capacitation agents (BWWCAP). A final study refined the combined use of permeating (0.68 or 1.35M glycerol) and non-permeating (0.2 or 0.3M sucrose) cryoprotectants. Spermatozoa were cryopreserved in liquid nitrogen vapour at rates of approximately -21°Cmin-1 (fast freeze) or -6.0°Cmin-1 (slow freeze). Post-thaw survival was highest with a combination of 0.2M sucrose and 0.68M glycerol and when these cryoprotectants were washed out with BWWCAP, regardless of whether spermatozoa were frozen using a fast (motility 14.2±4.7%; PI 20.7±2.0%) or slow (motility 12.0±2.7%; PI 22±4%) cryopreservation rate.

Measurement of testosterone and cortisol metabolites and luteinising hormone in captive southern hairy-nosed wombat (Lasiorhinus latifrons) urine.

This study reports the validation and use of enzyme immunoassays (EIA) to measure changes in plasma and urinary luteinizing hormone, testosterone metabolites (UTM) and cortisol metabolites (UCM) in captive southern hairy-nosed wombats (Lasiorhinus latifrons). GnRH agonist and ACTH agonist challenges were conducted to validate urinary testosterone (male wombat only) and cortisol (male and female wombats) EIAs. Following intra-muscular injection of 8-12µg buserelin (n=4 males), there was a significant increase in both plasma (P<0.001) and urinary testosterone concentrations (P<0.001) 60min and 21h after administration, respectively. Plasma LH levels were elevated (p<0.05) at 20min but there was no significant increase found in urinary LH concentrations after injection. Intra-muscular injection of Synacthen® Depot (250µg) (n=3 males, 3 females) resulted in a significant increase (p<0.05) in plasma cortisol secretion 15min and in urinary cortisol concentrations 3h post injection, respectively. Sex-related differences in cortisol secretion were also reported in this study. These findings indicate that (1) urinary LH might not be an appropriate index for describing the reproductive status in captive male L. latifrons, and (2) the UTM and UCM assays appear to be suitable for the assessment of the testicular steroidogenic capacity and the adrenocortical activity in captive southern hairy-nosed wombats, respectively.

An analysis of the complementarity of ICECAP-A and EQ-5D-3 L in an adult population of patients with knee pain.

BACKGROUND: The ICECAP measures potentially offer a broader assessment of quality of life and well-being, in comparison to measures routinely used in economic evaluation, such as the EQ-5D-3 L. This broader assessment may allow measurement of the full effects of an intervention or treatment. Previous research has indicated that the ICECAP-O (for older people) and EQ-5D-3 L measure provide complementary information. This paper aims to determine similar information for the ICECAP-A (for the entire adult population) in terms of whether the measure is a substitute or complement to the EQ-5D-3 L. METHODS: Data from the BEEP trial - a multi-centre, pragmatic, randomised controlled trial - were used. Spearman rank correlations and exploratory factor analytic methods were used to assess whether ICECAP-A and EQ-5D-3 L are measuring the same, or different, constructs. RESULTS: A correlation of 0.49 (p < 0.01) was found between the ICECAP-A tariff score and the EQ-5D-3 L index. Using the pooled items of the EQ-5D-3 L and the ICECAP-A a two factor solution was optimal, with the majority of EQ-5D-3 L items loading onto one factor and the majority of ICECAP-A items onto another. CONCLUSION: The results presented in this paper indicate that ICECAP-A and EQ-5D-3 L are measuring two different constructs and provide largely different, complementary information. Results showed a similarity to results presented by Davis et al. using the ICECAP-O. TRIAL REGISTRATION: ISRCTN 93634563.

A longitudinal assessment of the responsiveness of the ICECAP-A in a randomised controlled trial of a knee pain intervention.

PURPOSE: The ICECAP-A is a simple measure of capability well-being for use with the adult population. The descriptive system is made up of five key attributes: Stability, Attachment, Autonomy, Achievement and Enjoyment. Studies have begun to assess the psychometric properties of the measure, including the construct and content validity and feasibility for use. This is the first study to use longitudinal data to assess the responsiveness of the measure. METHODS: This responsiveness study was completed alongside a randomised controlled trial comparing three physiotherapy-led exercise interventions for older adults with knee pain attributable to osteoarthritis. Anchor-based methodologies were used to explore the relationship between change over time in ICECAP-A score (the target measure) and change over time in another measure (the anchor). Analyses were completed using the non-value-weighted and value-weighted ICECAP-A scores. The EQ-5D-3L was used as a comparator measure to contextualise change in the ICECAP-A. Effect sizes, standardised response means and t tests were used to quantify responsiveness. RESULTS: Small changes in the ICECAP-A scores were seen in response to underlying changes in patients' health-related quality of life, anxiety and depression. Non-weighted scores were slightly more responsive than value-weighted scores. ICECAP-A change was of comparable size to change in the EQ-5D-3L reference measure. CONCLUSION: This first analysis of the responsiveness using longitudinal data provides some positive evidence for the responsiveness of the ICECAP-A measure. There is a need for further research in those with low health and capability, and experiencing larger underlying changes in quality of life.

Faecal cortisol metabolites, body temperature, and behaviour of beef cattle exposed to a heat load.

Feedlot cattle are at times exposed to high environmental temperatures. Faecal cortisol metabolites were related to possible indicators of heat stress that could be measured under field conditions: respiratory dynamics (respiration rate), body surface temperature and adaptive behaviours, such as water consumption, posture (standing, lying), and activity (eating, drinking and rumination). Twelve (12) yearling Black Angus steers were divided into two treatment groups: a hot treatment (HOT; n = 6) and a thermoneutral-treatment (TN; n = 6) and individually housed in a climate-controlled facility at The University of Queensland, Australia. In the TN treatment, all animals were exposed to an ambient temperature of 20.34 ± 0.25 °C, relative humidity 71.51 ± 3.26% and Temperature humidity index (THI) 66.91 ± 0.33 throughout. In the HOT treatment group, environmental conditions were exposed to different climatic phases from thermoneutral to hot conditions, where they remained for 7 d, and then returned to TN conditions in the recovery period. The dry bulb ambient temperature (TA) and relative humidity (RH) in the pens of cattle in the HOT treatment were increased from 28 °C (daily maximum ambient temperature) and 45% RH at 0700 h to a daily maximum TA and RH of 35 °C (daily maximum ambient temperature) and 50% (THI 77) at 1100 h, which was maintained until 1600 h, after which it declined until it reached the baseline at 2000 h. In both treatments, there was a significant decrease in faecal cortisol metabolite concentration from the start to the end of the experiments they adapted to the experimental facility. The concentration of faecal cortisol metabolites was greater in the HOT treatment, compared to the TN treatment during the heat exposure period, but there was no difference in the transition or recovery periods. Respiration rate was greater in the HOT treatment during heat exposure, and it increased with ambient dry bulb temperature above 26 °C, the latter being the upper critical temperature. Although positive correlations were detected between faecal cortisol metabolites and body surface temperature measurements, particularly the shoulder and rump, as well as standing time, panting score and drinking, a stepwise regression found that faecal cortisol metabolites were only significantly correlated with one variable, respiration rate. It is concluded that respiration rate is the best indicator of the stress induced by hot conditions for cattle.

The effects of season and devil facial tumour disease on the reproductive physiology of the male Tasmanian devil (Sarcophilus harrisii).

Devil facial tumour disease (DFTD) is the cause of the rapid decline of wild Tasmanian devils. Female devils are seasonal breeders with births peaking during autumn (i.e. March) but the degree of reproductive seasonality in male devils is unknown. The objective of this study was to examine the potential effects of season and DFTD on reproductive function in male devils (n=55). Testicular (1.90±0.23 g) and epididymal (0.90±0.06 g) weights were maximal during autumn and spring (P<0.05), whereas prostate (3.71±0.74 g) and Cowper's gland (0.68±0.22; 0.52±0.21 g) weights peaked during autumn (P<0.001). The motility of spermatozoa from the cauda epididymides extracted post-mortem was similar (P>0.05) across season and disease state (31.5±13.1% total motility). Testicular and epididymal weights were no different between animals displaying late or early-stage DTFD signs or disease-free animals (P>0.1). The accessory sex glands were larger in late-stage DFTD animals than in animals with early-stage disease signs or which were disease-free (P<0.01) but effects of season on this result can't be excluded. Serum testosterone concentrations peaked during summer (0.25±0.18 ng mL(-1)) but values were not different from the preceding and subsequent seasons (P>0.05), nor influenced by disease stage (P>0.1). Seasonal and DFTD-related changes in serum cortisol concentrations were not evident (P>0.1). Male devil reproduction does not appear to be restricted by season nor inhibited by DFTD.

Development and evaluation of electroejaculation techniques in the Tasmanian devil (Sarcophilus harrisii).

Electroejaculation (EEJ) has been used successfully to collect samples suitable for genome resource banking from a variety of endangered wildlife species. Ejaculates can also be used to evaluate the reproductive potential of individuals and provide information on seminal characteristics to aid in the development of sperm cryopreservation techniques. Electroejaculation techniques used for marsupial and eutherian species were tested on Tasmanian devils (n=35). Spermic ejaculates were collected in 54% (19/35) of EEJ attempts. Spermic ejaculates were low in volume (3.9±6.5×10(2) µL, range 10-3000 µL) and contained low numbers of spermatozoa (3.3±7.8×10(3) spermatozoa per ejaculate, range 6-33000). The osmolality and pH of presumptive urine-free ejaculates were 389±130 mOsm kg(-1) (range 102-566) and 7.0±0.9 (range 6.0-8.0), respectively. Prostatic bodies were observed in 79% (26/33) of ejaculates. Episodic fluctuations in serum testosterone concentrations were not detected during the EEJ procedure (P>0.05). Increases observed in serum cortisol concentrations during EEJ were less (P<0.05) than those observed after an adrenalcorticotropic hormone challenge and diurnal variation suggested that cortisol concentrations are greater during the day than at night (P<0.05). This information can be used to provide range values for the future examination of basic endocrine responses and the adrenal-pituitary axis of this species. This study also demonstrated that spermatozoa-rich devil electroejaculates are more difficult to obtain and poorer in quality than those of other marsupials.

The reproductive cycle of the Tasmanian devil (Sarcophilus harrisii) and factors associated with reproductive success in captivity.

Numbers of wild Tasmanian devils are declining as a result of the fatal, transmissible Devil Facial Tumor Disease. A captive insurance population program has been initiated but current captive breeding rates are sub-optimal and therefore the goal of this project was to increase our understanding of the estrous cycle of the devil and elucidate potential causes of failed male-female pairings. Temporal patterns of fecal progestagen and corticosterone metabolite concentrations were examined for females (n=41) in three categories of reproductive status (successful: viable young, n=20 estrous cycles; unsuccessful: paired with a male but no young confirmed, n=44 estrous cycles; non-mated: no access to a male during estrus, n=8 estrous cycles) but substantial differences were not found. Females were more likely to produce pouch young if pairing with the male extended into late proestrus (P<0.05), thereby decreasing the time between pairing and presumed ovulation. The interval between the end of proestrous elevation in progestagen metabolite concentrations and the beginning of the luteal phase was 7.6±2.3 days in successful females. The length of the luteal phase in successful females was 12.5±1.4 days which was not different from unsuccessful or non-mated females (P>0.05). Unsuccessful females had 1-3 estrous cycles within a single year. Successful females were predominantly wild-caught (17/19, 90%) and most produced young following the first estrous cycle of the season (18/20, 90%). Unsuccessful females were predominantly captive born (20/27, 74%) in this study. It is possible that a proportion of females that do not produce pouch young achieve conception but the timing of reproductive failure continues to be elusive in this species.

The reproductive endocrinology and behavior of Vancouver Island marmot (Marmota vancouverensis).

The Vancouver Island marmot (Marmota vancouverensis; VIM) is one of North America's most endangered species with fewer than 150 individuals remaining in the wild. A captive breeding program was established across four facilities in Canada as an insurance population and source of animals for reintroduction to the wild. The purpose of this study was to gather information about the basic reproductive biology and behavior of this species, which is essential to improve captive breeding programs. Regular fecal samples were obtained from adult female (n = 14) and male (n = 10) marmots, 2 years of age and older, over 1-3 breeding seasons (2-3 months duration posthibernation) for steroid hormone analysis. Enzyme immunoassays were validated for quantifying fecal testosterone metabolite concentrations for males, and fecal estrogen and progesterone metabolite concentrations for females. Results indicated that fecal progesterone metabolite concentrations can be used to monitor ovulation and pregnancy. Behavioral monitoring through infrared video surveillance was conducted in four breeding pairs over a 2-year period (n = 7 behavioral profiles). Breeding behaviors correlated strongly with changes in reproductive endocrine profiles. A high frequency of play behavior or "wrestling" was observed in conjunction with breeding activity before an elevation in progesterone metabolite concentrations. Impending parturition was associated with increased aggression and exclusion of the male from the maternal nestbox as well as an increase in nesting activity. Observational data combined with hormonal analysis suggest that female VIMs are induced ovulators and that multiple breeding attempts may be required for ovulation and conception. Gestation appears to be approximately 34 days from peak breeding activity (32 days from estimated ovulation). Fecal testosterone concentrations suggest that testicular activity is seasonal with the reproductive activity occurring immediately posthibernation. Monitoring breeding behavior is a useful means of indicating estrus, conception and pregnancy, which can also be supported by the hormonal analysis of daily fecal samples of individual animals.

Systematic review of the use of translated patient-reported outcome measures in cancer trials.

BACKGROUND: Patient-reported outcomes (PROs) are used in clinical trials to assess the effectiveness and tolerability of interventions. Inclusion of participants from different ethnic backgrounds is essential for generalisability of cancer trial results. PRO data collection should include appropriately translated patient-reported outcome measures (PROMs) to minimise missing data and sample attrition. METHODS: Protocols and/or publications from cancer clinical trials using a PRO endpoint and registered on the National Institute for Health Research Portfolio were systematically reviewed for information on recruitment, inclusion of ethnicity data, and use of appropriately translated PROMs. Semi-structured interviews were conducted with key stakeholders to explore barriers and facilitators for optimal PRO trial design, diverse recruitment and reporting, and use of appropriately translated PROMs. RESULTS: Eighty-four trials met the inclusion criteria, only 14 (17%) (n = 4754) reported ethnic group data, and ethnic group recruitment was low, 611 (13%). Although 8 (57%) studies were multi-centred and multi-national, none reported using translated PROMs, although available for 7 (88%) of the studies. Interviews with 44 international stakeholders identified a number of perceived barriers to ethnically diverse recruitment including diverse participant engagement, relevance of ethnicity to research question, prominence of PROs, and need to minimise investigator burden. Stakeholders had differing opinions on the use of translated PROMs, the impact of trial designs, and recruitment strategies on diverse recruitment. Facilitators of inclusive research were described and examples of good practice identified. CONCLUSIONS: Greater transparency is required when PROs are used as primary or secondary outcomes in clinical trials. Protocols and publications should demonstrate that recruitment was accessible to diverse populations and facilitated by trial design, recruitment strategies, and appropriate PROM usage. The use of translated PROMs should be made explicit when used in cancer clinical trials.

Testosterone secretion, testicular histology and the cryopreservation of cauda epididymidal spermatozoa in the common ringtail possum (Pseudocheirus peregrinus).

The present study reports novel aspects of the reproductive biology of the male common ringtail possum (Pseudocheirus peregrinus). Plasma testosterone was measured through a stimulation test using the gonadotrophin-releasing hormone agonist, buserelin. Following intra-muscular administration of buserelin, there was an increase (P<0.05) in testosterone concentration in the peripheral circulation 4 h later. Quantitative testicular histology of this species was described for the first time. Eight stages of the seminiferous epithelium cycle were identified in 10 possums and their relative frequency determined. Spermatozoa were recovered from the cauda epididymides of hemi-castrated possums and cryopreservation conducted in straws (6 degrees C min(-1)) using final glycerol concentrations ranging between 2 and 20% in Tris-citrate egg yolk extender (v/v). Frozen straws were thawed and post-thaw motility, rate of motility, the percentage of live-dead spermatozoa and the percentage of sperm with swollen decondensed nuclei recorded. Similar to other marsupial sperm, common ringtail possum cauda epididymidal spermatozoa required high levels of glycerol (10-16%) in order to maintain post-thaw viability.

Studies of male reproduction in captive African wild dogs (Lycaon pictus).

Implementation of assisted breeding in the captive African wild dog is restricted by a current lack of knowledge on their reproductive physiology and the apparent difficulty of effectively manipulating the complex social dynamic of the pack in order to conduct reproductive procedures. In this study, we describe protocols for the safe and repeated capture and restraint of the African wild dog (n=7) as well as techniques for assessment of male reproductive function, semen collection and preservation. In a serendipitous finding, captive African wild dogs appeared to display significant seasonal change in male reproduction. Testicular volume and tone, spermatorrhea and the ability to collect semen by electroejaculation all increased significantly during late summer and then subsequently declined by early spring. While there were no detectable seasonal changes in testosterone concentration in the population as whole, the alpha-dominant male in both years of the study, had a highly elevated testosterone concentration compared to subordinate males. Semen collection by electroejaculation during the late summer was most effective in peri-pubertal males (15 months) when all seven electroejaculates were of adequate quality for cryopreservation. In the second breeding season (27 months), there were numerous changes in the pack hierarchy and electroejaculation was not as successful (3/7). The characteristics of electroejaculated semen collected in the breeding season are described for seven animals including the first descriptions and incidence of sperm abnormalities in the species. Semen (n=7) was frozen using a Tris-citrate fructose buffer and final egg yolk and glycerol concentration of 20% and 4%, respectively. Sperm were loaded into 0.25 mL straws, frozen in liquid nitrogen vapor and then thawed at 37 degrees C. Initial post-thaw survival of spermatozoa was encouraging (% motile: 31.8+/-5.8%; rate: 2.8+/-0.3; % intact plasma membranes: 33.4+/-5.3% and the % of damaged acrosomes: 4.4+/-1.5%) but following 2 h incubation at 37 degrees C, post-thaw survival declined markedly.

Seasonality and breeding success of captive and wild Tasmanian devils (Sarcophilus harrisii).

The synchrony and timing of reproductive events are crucially important factors to maximize individual and offspring survival, especially in seasonal environments. To increase our understanding of the physiological basis of seasonality and the influence of associated environmental factors (maximum temperature, day length and rate of day length change associated with different latitudes) on reproduction in Tasmanian devils, we reviewed records and research data from captive facilities throughout Australia in comparison to those from a wild population study (1974-1987). Overall, breeding activity began 2 weeks earlier in the captive than the wild population (week 5.7 ± 0.6 versus week 7.7 ± 0.5 for devils entering into estrus during the first two week phase; n = 24 and n = 23 respectively). If the timing of reproductive activity is considered against absolute day length rather than date, both the captive and wild populations displayed similar distributions (12.9 ± 0.7 h versus 13.0 ± 0.7 h respectively; P < 0.01) confirming day length as a proximal cue involved in eliciting a physiological response to trigger seasonal reproductive activity regardless of location. Wild devils had a higher breeding success (75%; n = 169 versus 43%; n = 115) and larger litter size (3.4 ± 0.9 versus 2.8 ± 1.1 joeys per litter) than captive devils (P < 0.05). Mean maximum temperature at the onset of reproductive activity (P < 0.05) was higher for the captive than the wild population (28.1 ± 4.0 °C versus 22.3 ± 2.7 °C respectively). The drivers for reproductive success in captive Tasmanian devils are likely multifactorial, but our results suggest that elevated temperatures associated with shifts in breeding activity and geographical location should be examined further.

First report of a spermatic granuloma and varicocele in a marsupial: A Koala (Phascolarctos cinereus) Case Study.

This study reports the first documented clinical case of a spermatic granuloma and varicocele in a marsupial. Initial clinical presentation included gross morphological changes in the left scrotal cord, epididymis and testis. Ultrasonography of the scrotum and spermatic cord, and gross and histopathological examination after hemicastration, confirmed the condition as a spermatic granuloma affecting the left caput epididymis, with a varicocele in the left proximal spermatic cord, which was causing azoospermia and infertility. Semen quality and serum testosterone secretion following a GnRH challenge was assessed prior to, and following surgery. After hemi-castration, an increase in androgen secretion to within normal reference ranges for the koala was observed with a subsequent increase in semen production and sperm quality resulting in the sire of a pouch young, 12months later.

The use of qualitative methods to inform Delphi surveys in core outcome set development.

BACKGROUND: Core outcome sets (COS) help to minimise bias in trials and facilitate evidence synthesis. Delphi surveys are increasingly being used as part of a wider process to reach consensus about what outcomes should be included in a COS. Qualitative research can be used to inform the development of Delphi surveys. This is an advance in the field of COS development and one which is potentially valuable; however, little guidance exists for COS developers on how best to use qualitative methods and what the challenges are. This paper aims to provide early guidance on the potential role and contribution of qualitative research in this area. We hope the ideas we present will be challenged, critiqued and built upon by others exploring the role of qualitative research in COS development. This paper draws upon the experiences of using qualitative methods in the pre-Delphi stage of the development of three different COS. Using these studies as examples, we identify some of the ways that qualitative research might contribute to COS development, the challenges in using such methods and areas where future research is required. RESULTS: Qualitative research can help to identify what outcomes are important to stakeholders; facilitate understanding of why some outcomes may be more important than others, determine the scope of outcomes; identify appropriate language for use in the Delphi survey and inform comparisons between stakeholder data and other sources, such as systematic reviews. Developers need to consider a number of methodological points when using qualitative research: specifically, which stakeholders to involve, how to sample participants, which data collection methods are most appropriate, how to consider outcomes with stakeholders and how to analyse these data. A number of areas for future research are identified. CONCLUSIONS: Qualitative research has the potential to increase the research community's confidence in COS, although this will be dependent upon using rigorous and appropriate methodology. We have begun to identify some issues for COS developers to consider in using qualitative methods to inform the development of Delphi surveys in this article.

Effect of age and presence of perches during rearing on tonic immobility fear reactions of broiler breeder pullets.

Broiler breeders were housed in pens in an all-litter house with an 8-h photoperiod from hatching to 20 wk of age. They were reared either in the presence or absence of perches (7 cm per bird) and on either a 14% or a 17% CP diet. The duration of the tonic immobility (TI) fear response was measured in 15- and in 20-wk-old birds; each pullet was tested individually and once only. There was an apparent age-related increase in the duration of TI, which was attenuated by the provision of perches during rearing. The results are discussed in terms of maturational and environmental influences on the development and alleviation of fear.