Evidence for contrasting roles for prolactin in eusocial naked mole-rats, Heterocephalus glaber and Damaraland mole-rats, Fukomys damarensis

Elevated prolactin (PRL) has been associated with the expression of social and cooperative behaviours in a number of vertebrate species, as well as suppression of reproduction. As social mole-rats exhibit both of these traits, PRL is a prime candidate in mediating their social phenotype. While naked and Damaraland mole-rats (NMRs and DMRs) have evolved eusociality independently within their family, both species exhibit an extreme skew in lifetime reproductive success, with breeding restricted to a single female and one or two males. Non-breeding NMRs of both sexes are physiologically inhibited from reproducing, while in DMRs only the non-breeding females are physiologically suppressed. Newly emerging work has implicated the dopamine system and PRL as a component in socially induced reproductive suppression and eusociality in NMR, but the DMR remains unstudied in this context. To investigate evolutionary convergence in the role of PRL in shaping African mole-rat eusociality, we determined plasma PRL concentrations in breeders and non-breeders of both sexes, comparing DMRs with NMRs. Among samples from non-breeding NMRs 80% had detectable plasma PRL concentrations. As a benchmark, these often (37%) exceeding those considered clinically hyperprolactinaemic (25 ng ml−1) in humans: mean ± s.e.m.: 34.81 ± 5.87 ngml−1; range 0.00–330.30 ng ml−1. Conversely, 85% of non-breeding DMR samples had undetectable values and none had concentrations above 25 ng ml−1: 0.71 ± 0.38 ng ml−1; 0.00–23.87 ngml−1. Breeders in both species had the expected variance in plasma PRL concentrations as part of normal reproductive function, with lactating queens having significantly higher values. These results suggest that while elevated PRL in non-breeders is implicated in NMR eusociality, this may not be the case in DMRs, and suggests a lack of evolutionary convergence in the proximate control of the social phenotype in these mole-rats.SARCHI Chair to NCB (Grant N64756)http://rsbl.royalsocietypublishing2019-05-01hj2018Anatomy and PhysiologyMammal Research InstituteZoology and Entomolog

Behavioural inventory of the giraffe (<i>Giraffa camelopardalis</i>)

Background Numerous factors like continuous habitat reduction or fragmentation for free-ranging giraffes (Giraffa camelopardalis) as well as e.g. suboptimal housing conditions for animals in captivity might lead to behavioural alterations as part of the overall adaptation process to the changing living conditions. In order to facilitate current and future studies on giraffe behaviour, a comprehensive ethogram was compiled based on existing literature, as well as observations on giraffes in the wild (Hwange National Park, Zimbabwe; Entabeni Game Reserve, South Africa), and in captivity (National Zoological Gardens of South Africa, Pretoria). Findings The resulting ethogram lists 65 different behavioural patterns, which were described and grouped into seven categories: General activities, Abnormal repetitive behaviours, General interactions, Bull-Cow behaviour, Bull-Bull behaviour, Cow-Bull behaviour, Maternal behaviours, and Interactions by calves. The behaviours were further described regarding a presumed purpose, particularly with respect to social interactions and sexual behaviour. Contradictory descriptions from previous studies were considered and discussed in comparison with our own observations. Conclusions This ethogram provides a basis for current and future studies by suggesting a terminology which can be used for harmonizing behavioural observations, thus helping to facilitate comparability of future results. Subsequently, a better understanding of the behavioural ecology of giraffes in the wild as well as in captivity could aid future conservation efforts

A Bayesian latent class model to estimate the accuracy of pregnancy diagnosis by transrectal ultrasonography and laboratory detection of pregnancy-associated glycoproteins in dairy cows

Accurate diagnosis of pregnancy is an essential component of an effective reproductive management plan for dairy cattle. Indirect methods of pregnancy detection can be performed soon after breeding and offer an advantage over traditional direct methods in not requiring an experienced veterinarian and having potential for automation. The objective of this study was to estimate the sensitivity and specificity of pregnancy-associated glycoprotein (PAG) detection ELISA and transrectal ultrasound (TRUS) in dairy cows of South Africa using a Bayesian latent class approach. Commercial dairy cattle from the five important dairy regions in South Africa were enrolled in a short-term prospective cohort study. Cattle were examined at 28–35 days after artificial insemination (AI) and then followed up 14 days later. At both sampling times, TRUS was performed to detect pregnancy and commercially available PAG detection ELISAs were performed on collected serum and milk. A total of 1236 cows were sampled and 1006 had complete test information for use in the Bayesian latent class model. The estimated sensitivity (95% probability interval) and specificity for PAG detection serum ELISA were 99.4% (98.5, 99.9) and 97.4% (94.7, 99.2), respectively. The estimated sensitivity and specificity for PAG detection milk ELISA were 99.2% (98.2, 99.8) and 93.4% (89.7, 96.1), respectively. Sensitivity of veterinarian performed TRUS at 28–35 days post-AI varied between 77.8% and 90.5% and specificity varied between 94.7% and 99.8%. In summary, indirect detection of pregnancy using PAG ELISA is an accurate method for use in dairy cattle. The method is descriptively more sensitive than veterinarian-performed TRUS and therefore could be an economically viable addition to a reproductive management plan

Concentrations of faecal glucocorticoid metabolites in South African Mutton Merino sheep fed various sources of roughage

The objective of this study was to determine whether various sources of roughage in nutritionally balanced feedlot diets would evoke a stress response in sheep. A nutritional stress response test was performed on 20 ewes, randomly divided into four treatment groups with 5 ewes per group. Ewes were kept individually in metabolic crates for thirty days. Rations were balanced nutritionally, and various sources of roughage were included; T1: alfalfa hay, T2: maize stover, T3: soya hulls; and T4: Eragrostis tef. Faeces was removed manually from the caudal rectum of each ewe at 05h00 and 19h00. Samples were frozen at -20°C until analyses. A total of 520 faecal samples were analysed to determine the concentrations of faecal glucocorticoid metabolites (fGCM) by enzyme immunoassay (EIA). These concentrations were expressed as mass/g dry weight (ng/g DW). Baseline values were determined, and those greater than the mean plus 2 standard deviations were removed. Baseline values were compared between the groups with one-way ANOVA analysis. The average concentrations of fGCM were 178.77 ± 21.7 in the morning and 183.2 ± 14.4 in the evening. Sheep fed T4 had significantly higher fGCM concentrations in both morning (302.0 ± 86.1 ng/g DW) and evening (237.0 ± 48.1 ng/g DW) compared with the other treatment groups. Thus, fGCM concentrations in sheep were related to the source of roughage, and Eragrostis tef caused a stress response. Keywords: animal welfare, cortisol, dietary fibre, selective grazer, stress respons

Normalized Difference Vegetation Index, temperature, and age affect faecal thyroid hormone concentrations in free-ranging African elephants.

Conservation biologists can use hormone measurements to assess animals’ welfare, reproductive state, susceptibility to stressors, as well as energy expenditure. Quantifying hormone concentrations from faecal samples is particularly advantageous as samples can be collected without disturbing animals’ behaviour. In order for an endocrine marker to be useful for wildlife managers, we need to understand how extrinsic and intrinsic factors affect hormone concentrations in free-ranging animal populations. Thyroid hormones are linked to basal metabolic rate and energy expenditure. Previous research demonstrated that triiodothyronine (T3) can be measured successfully in faecal matter of African elephants, Loxodonta africana. However, to our knowledge, research into factors affecting changes in elephant T3 levels has only been carried out in captive elephants so far. Thus, we present the first study of faecal T3 metabolite (mT3) concentrations of a large population of free-ranging African elephants. Over 15 months, we collected faecal samples from identified (n=43 samples) and unidentified (n=145 samples) individuals in Madikwe Game Reserve, South Africa. We investigated whether vegetative productivity (NDVI) in interaction with mean monthly temperature, age, and sex affected mT3 concentrations. We found a significant negative interaction effect of NDVI and temperature. Increasing NDVI was related to higher concentrations of mT3, but increasing temperature was related to a decrease in mT3 concentrations in individually identified and unidentified elephants. In unidentified individuals, juvenile elephants had significantly higher mT3 concentrations compared to adult elephants. Faecal T3 can successfully be quantified in samples from free-ranging elephant populations and thus provides insight into energy expenditure in large herbivores

Social dominance does not affect semen quality in African wild dogs (Lycaon pictus)

Sperm banking and artificial insemination could benefit conservation of endangered African wild dogs (AWD). However, it is not clear whether their strict dominance hierarchy causes subfertility in subdominant males that typically do not breed. Our study investigated the effect of dominance on male reproductive parameters including: faecal glucocorticoids (fGCMs) and androgens (fAMs), testis and prostate volume, preputial gland size, semen collection success, and the number, motility, morphology, viability, acrosome integrity (PSA-FITC), and DNA integrity (TUNEL) of spermatozoa collected by electroejaculation. Samples were obtained from n=12 captive AWDs (4 US packs) in the pre-breeding season and n=28 captive AWDs (n=11 from 4 US packs; n=17 from 3 Namibian packs) in the breeding season. Male hierarchy was clearly determined by behavioural observations in all but 1 Namibian pack. Data were grouped by dominance status and means compared by ANOVA or t-test. P≤0.05 was significant. In the pre-breeding season, there was no significant difference in body weight, fGCMs, fAMs, or prostate and testis volume between dominance groups. Semen was successfully collected from all alphas but only half the subdominants; with urine contamination negatively associated with dominance. Sperm quality was low (17.3 ± 10.2% total motility, 12.8 ± 8.5% progressive motility, 27.4 ± 11.5 x 106 ejaculated spermatozoa, 40.6 ± 9.8% normal morphology, 63.1 ± 5.1% viability, 72.6 ± 5.2% acrosome integrity) with no difference observed in any parameter except progressive motility and normal sperm morphology; which were significantly lower in subdominants (27.7 ± 16.8% vs. 0.0 ± 0.0% and 59.8 ± 13.0% vs. 21.4 ± 5.7%). From pre-breeding to breeding season, testis and prostate volume increased significantly; particularly in beta and gamma males respectively. Prostate volume was higher in alpha than beta males (16.0 ± 6.4 cm3 vs. 5.7 ± 1.4 cm3), but testis volume, body weight, fAMs and fGCMs did not differ between dominance groups (12.0 ± 0.9 cm3, 28.5 ± 0.8 kg, 0.51 ± 0.07 µg/g dry weight - DW, 30.6 ± 2.3 ng/g DW). Semen was successfully collected from 75% of males; with reduced urine contamination. Collection success, urine contamination and preputial gland size were not associated with dominance. Sperm quality improved with significantly greater number, viability, and total motility. However, sperm quality did not differ between dominance groups (47.4 ± 6.7% total motility, 30.5 ± 5.8% progressive motility, 32.3 ± 9.2 x 106 ejaculated spermatozoa, 50.9 ± 5.2% normal morphology, 74.4 ± 4.2% viability, 85.6 ± 3.0% acrosome integrity, 99.7 ± 0.1% DNA integrity). In conclusion, subdominant males are at higher risk of urine contamination and have lower sperm motility and normal morphology when semen is collected in the pre-breeding season. However, their semen is of similar quality to dominant males in the breeding season, indicating that reproductive suppression of subdominant males is only behavioural. Thus, AWD males of all social ranks in the breeding season are suitable candidates for sperm banking

Dog Appeasing Pheromone prevents the testosterone surge, and may reduce contact-dominance and active-submission behaviours after interventions in captive African wild dogs (Lycaon pictus)

African wild dogs (Lycaon pictus) have a complex hierarchical social structure that can lead to aggression; resulting in morbidity and mortality of individuals separated from the pack, or during new pack formation. Aggression and stress might be attenuated by Dog Appeasing Pheromone (DAP). Our placebo-controlled, double-blinded study showed an increase in faecal androgen concentrations within the pack after temporary separation, immobilisation and reintroduction of placebo treated animals, but not for DAP treated individuals. Moreover, DAP treated packs tended to show lower rates of contact-dominance and activesubmission behaviour, but higher rates of non-contact dominance behaviour. Faecal corticosteroid levels rose after intervention in both treatments, probably due to immobilisation-related stress. These preliminary findings suggest that DAP could be a useful management tool to reduce hormones and behaviours potentially leading to aggression in captive African wild dogs

Non-invasive measurement of adrenocortical activity in a myrmecophageous mammal, the bat-eared fox

Measuring physiological stress reactions through the quantification of plasma cortisol often involves physically restraint, which acts as a stressor itself. Here, we present the validation of a non-invasive method for assessing adrenocortical activity as an indicator of stress in the bat-eared fox (Otocyon megalotis). By conducting an adrenocorticotropic hormone (ACTH) challenge, we examined the suitability of three enzyme immunoassays (EIAs) detecting 11,17 dioxoandrostanes (11,17-DOA) as well as faecal glucocorticoid metabolites (fGCM) with a 5β-3α-ol-11-one (3α,11oxo-CM), or 11,17,21- trihydroxy-4-ene-20-one structure (cortisol), respectively, for monitoring stress-related physiological responses in male and female bat-eared foxes. Our results suggest that the cortisol EIA seems most suitable for measuring fGCMs in this myrmecophageous mammal. Using the cortisol EIA, we compared fGCM concentrations of three populations of foxes. Only one population experienced a sudden change in social environment and the assay appeared to effectively detect the resulting increase in stress hormone levels. Therefore, the identified EIA is effective at detecting intraspecific variation in fGCM levels, and hence a useful tool to evaluate physiological stress responses in this species.Thuthuka grant (#TTK1206041007) from the National Research Foundation.http://www.tandfonline.com/loi/tafz202017-02-28hb2016Anatomy and PhysiologyMammal Research InstituteZoology and Entomolog

Putative drivers of adrenocortical activity in captive African lesser bushbaby ( Galago moholi )

In seasonal breeders, periods of reproductive activity often coincide with high levels of glucocorticoids. We studied seven male and seven female African lesser bushbabies (Galago moholi A. Smith, 1836) over two mating periods via noninvasive faecal hormone metabolite monitoring to investigate the relationship between reproductive and adrenocortical hormone activity. We used linear mixed-effect models to investigate the effect of physiological (endocrine) variables on faecal glucocorticoid metabolite concentrations. Our results indicate faecal androgen (males) and progestagen metabolite (females) concentrations as the variables best able to explain variability in faecal glucocorticoid metabolite concentrations. However, the models explained only a fraction (26% and 12%, respectively) of the observed variability and graphical analysis suggests a biologically relevant difference in faecal glucocorticoid metabolite concentrations between captive and free-ranging animals during nonreproductive periods. Thus, captivity may have affected glucocorticoid output in our focal animals, potentially weakening the expected relationship between reproductive activity and faecal glucocorticoid metabolite variability. Due to the ease of faecal and observational sample collection, a large number of studies monitoring adrenocortical activity in wildlife are conducted using only captive settings, with inferences unquestioned when applied to free-ranging scenarios. Our study cautions against this practice, as particular housing or management conditions may influence the pattern of adrenocortical activity