Evaluation of adrenocortical function in Florida manatees (Trichechus manatus latirostris)

The study objectives were to determine the predominant manatee glucocorticoid; validate assays to measure this glucocorticoid and adrenocorticotropic hormone (ACTH); determine diagnostic thresholds to distinguish physiological vs. pathological concentrations; identify differences associated with sex, age class, female reproductive status, capture time, and lactate; and determine the best methods for manatee biologists and clinicians to diagnose stress. Cortisol is the predominant manatee glucocorticoid. IMMULITE 1000 assays for cortisol and ACTH were validated. Precision yielded intra‐ and inter‐assay coefficients of variation for serum cortisol: ≤23.5 and ≤16.7%; and ACTH: ≤6.9 and ≤8.5%. Accuracy resulted in a mean adjusted R²≥0.87 for serum cortisol and ≥0.96 for ACTH. Assay analytical sensitivities for cortisol (0.1 µg/dl) and ACTH (10.0 pg/ml) were verified. Methods were highly correlated with another IMMULITE 1000 for serum cortisol (r=0.97) and ACTH (r=0.98). There was no significant variation in cortisol or ACTH with sex or age class and no correlation with female progesterone concentrations. Cortisol concentrations were highest in unhealthy manatees, chronically stressed by disease or injury. ACTH was greatest in healthy free‐ranging or short‐term rehabilitating individuals, peracutely stressed by capture and handling. Cortisol concentrations ≥1.0 µg/dl were diagnostic of chronic stress; ACTH concentrations ≥87.5 pg/ml were diagnostic of peracute stress. In healthy long‐term captive manatees, cortisol (0.4±0.2 µg/dl) and ACTH (47.7±15.9 pg/ml) concentrations were lower than healthy free‐ranging, short‐term rehabilitated or unhealthy manatees. Capture time was not significantly correlated with cortisol; ACTH correlation was borderline significant. Cortisol and ACTH were positively correlated with lactate. Zoo Biol 30:17–31, 2011. © 2010 Wiley‐Liss, Inc.     

Using human sequencing to guide craniofacial research

A recent convergence of technological innovations has re‐energized the ability to apply genetics to research in human craniofacial development. Next‐generation exome and whole genome sequencing have significantly dropped in price, making it relatively trivial to sequence and analyze patients and families with congenital craniofacial anomalies. A concurrent revolution in genome editing with the use of the CRISPR‐Cas9 system enables the rapid generation of animal models, including mouse, which can precisely recapitulate human variants. Here, we summarize the choices currently available to the research community. We illustrate this approach with the study of a family with a novel craniofacial syndrome with dominant inheritance pattern. The genomic analysis suggested a causal variant in AMOTL1 which we modeled in mice. We also made a novel deletion allele of Amotl1. Our results indicate that Amotl1 is not required in the mouse for survival to weaning. Mice carrying the variant identified in the human sequencing studies, however, do not survive to weaning in normal ratios. The cause of death is not understood for these mice complicating our conclusions about the pathogenicity in the index patient. Thus, we highlight some of the powerful opportunities and confounding factors confronting current craniofacial genetic research.     

Multiple paternity in the European hedgehog

Previous observations have suggested that European hedgehogs Erinaceus europaeus are promiscuous, although there have been no direct studies of the mating system to confirm this. In this study, the genetic mating system of the European hedgehog was assessed by analysing mother–offspring groups using six polymorphic microsatellite loci. Evidence of multiple paternity was found in two of five litters analysed and therefore demonstrates the occurrence of polyandry in wild populations of hedgehogs.     

Generation and characterization of a Ddx4-iCre transgenic line for deletion in the germline beginning at genital ridge colonization

Germline-specific Cre lines are useful for analyses of primordial germ cell, spermatogonial and oogonial development, but also for whole-body deletions when transmitted through subsequent generations. Several germ cell specific Cre mouse strains exist, with various degrees of specificity, efficiency, and temporal activation. Here, we describe the CRISPR/Cas9 targeted insertion of an improved Cre (iCre) sequence in-frame at the 3′ end of the Ddx4 locus to generate the Ddx4-P2A-iCre allele. Our functional assessment of this new allele, designated Ddx4^iCreJoBo, reveals that Cre activity begins in PGCs from at least E10.5, and that it achieves higher efficiency for early gonadal (E10.5–12.5) germline deletion when compared to the inducible Oct4^CreERT2 line. We found the Ddx4^iCreJoBo allele to be hypomorphic for Ddx4 expression and homozygous males, but not females, were infertile. Using two reporter lines (R26R^LacZ and R26R^tdTomato) and a floxed gene of interest (Cripto^flox) we found ectopic activity in multiple organs; global recombination (a common feature of germline Cre alleles) varies from 10 to 100%, depending on the particular floxed allele. There is a strong maternal effect, and therefore it is preferable for Ddx4^iCreJoBo to be inherited from the male parent if ubiquitous deletion is not desired. With these limitations considered, we describe the Ddx4^iCreJoBo line as useful for germline studies in which early gonadal deletion is required.     

Current and future impacts of nest predation and nest-site competition by invasive eastern grey squirrels Sciurus carolinensis on European birds

1. The eastern grey squirrel (hereafter ‘grey squirrel’) is considered one of the most damaging invasive alien species in Europe, with negative effects on native ecosystems. Despite it being widely perceived as a significant predator of bird eggs and chicks and as a competitor for nest sites, evaluation of the grey squirrel’s impact on European bird populations has been hindered by limited empirical data.
2. The aim was to review the incidence of grey squirrels as nest predators of and nest-site competitors with European birds, and to use this information to identify species at potential risk of negative effects from within the grey squirrel’s expanding range in continental Europe.
3. A comprehensive literature review was conducted, and the data were used alongside additional new data, to assess nest predation and competition by grey squirrels in their current European range. Bird species were grouped by nest-site type, which was used to predict the impact on similar species groups in regions of continental Europe predicted to be colonised by grey squirrels in the current century.
4. Camera-monitoring and field evidence for 12 bird species and 12420 nests in Britain showed that grey squirrels rarely depredated eggs or chicks, affecting just 0.5% of nests. Nest-site competition was also minor, with grey squirrels occupying 0.8% of 122 small tree cavities and 14% of 57 larger cavities. At least 69 bird species in continental Europe could be exposed to potential nest predation or competition from expanding grey squirrel populations within the current century, but population-level effects currently appear to be unlikely.
5. Current evidence shows that grey squirrels are unlikely to be significant predators of or competitors with nesting birds in their present or projected range in Europe. However, further studies of more species in different regions would be valuable, particularly in urban and suburban habitats.

     

Cope's rule and the evolution of body size in Pinnipedimorpha (Mammalia: Carnivora)

Cope's rule describes the evolutionary trend for animal lineages to increase in body size over time. In this study, we tested the validity of Cope's rule for a marine mammal clade, the Pinnipedimorpha, which includes the extinct Desmatophocidae, and extant Phocidae (earless seals), Otariidae (fur seals and sea lions), and Odobenidae (walruses). We tested for the presence of Cope's rule by compiling a large dataset of body size data for extant and fossil pinnipeds and then examined how body size evolved through time. We found that there was a positive relationship between geologic age and body size. However, this trend is the result of differences between early assemblages of small-bodied pinnipeds (Oligocene to early Miocene) and later assemblages (middle Miocene to Pliocene) for which species exhibited greater size diversity. No significant differences were found between the number of increases or decreases in body size within Pinnipedimorpha or within specific pinniped clades. This suggests that the pinniped body size increase was driven by passive diversification into vacant niche space, with the common ancestor of Pinnipedimorpha occurring near the minimum adult body size possible for a marine mammal. Based upon the above results, the evolutionary history of pinnipeds does not follow Cope's rule.     

GROWTH LAYER PATTERNS IN TEETH FROM THE FRANCISCANA, PONTOPORIA BLAINVILLEI: DEVELOPING A MODEL FOR PRECISION IN AGE ESTIMATION

A model for estimating age from teeth of the franciscana, Pontoporia blainvillei , was developed using teeth from 72 specimens taken incidentally in fishing nets in Brazil and Uruguay. The goal of the model was to allow for accurate and precise age estimates for a larger sample of specimens. Growthlayer characteristics were selected on the basis of similarities in growth layers among various species of cetaceans, including those whose growth layers have been calibrated for time, and on length groups in the franciscana, which calves seasonally. Growth-layer patterns, including relative size and appearance, in both dentine and cementum did not vary by sex but did vary by the orientation of the tooth when it was sectioned. The model developed proved useful in providing guidelines for consistency in age estimation for biological studies of the larger sample of animals. We recommend the development of this type of model for other species to facilitate standardizing age estimation.     

Evolution of the secretoglobins: a genomic and proteomic view

Mouse salivary androgen-binding protein (ABP) is a member of the newly erected secretoglobin family, no member of which has yet been assigned an indisputable function. We have suggested a role for ABP in mate selection behaviour and sexual isolation. Although this has been a particularly attractive hypothesis given the evidence for strong positive selection on its alpha subunit gene, Abpa , we have held out the possibility that there might be an as-yet-undiscovered primary function for ABP. This is particularly important in light of its membership in the secretoglobin family, and we are pursuing the broader issue of shared functions of the secretoglobins with genomic and bioinformatic studies. Here we present as complete a comparison as possible of the secretoglobins in the genomes of three species of mammals: mouse, rat and human, and we compare the protein sequences and their potential evolutionary relationships. We suggest that the secretoglobins can be divided into at least five families. In rodent and human genomes, these gene families are found in two main clusters that are syntenic between rat and mouse. Humans have only the three families that are found within the uteroglobin/clara cluster, because no ABP-containing secretoglobin cluster has yet been identified.  © 2005 The Linnean Society of London, Biological Journal of the Linnean Society , 2005, 84 , 493–501.     

Scaling of sensorimotor control in terrestrial mammals

Sensorimotor control is greatly affected by two factors—the time it takes for an animal to sense and respond to stimuli (responsiveness), and the ability of an animal to distinguish between sensory stimuli and generate graded muscle forces (resolution). Here, we demonstrate that anatomical limitations force a necessary trade-off between responsiveness and resolution with increases in animal size. To determine whether responsiveness is prioritized over resolution, or resolution over responsiveness, we studied how size influences the physiological mechanisms underlying sensorimotor control. Using both new electrophysiological experiments and existing data, we determined the maximum axonal conduction velocity (CV) in animals ranging in size from shrews to elephants. Over the 100-fold increase in leg length, CV was nearly constant, increasing proportionally with mass to the 0.04 power. As a consequence, larger animals are burdened with relatively long physiological delays, which may have broad implications for their behaviour, ecology and evolution, including constraining agility and requiring prediction to help control movements.