Dynamic Fracture Resistance under Plane Strain Conditions of High-Density Polyethylene Nanoclay Composites.

Polymer nanoclay composites have received significant attention due to their substantially enhanced mechanical, thermal and barrier properties. However, the effect of these nanoclays on the dynamic fracture resistance of a polymer matrix during fast fracture events has not been documented. In this study, the effect of nanoclay addition on the rapid crack propagation (RCP) resistance of high-density polyethylene (HDPE) was investigated through the high-speed double torsion test. Results showed that the addition of 1, 3, and 5% of nanoclays improved the dynamic fracture resistance under the plane strain conditions (Gd1) of HDPE up to 65%. An increase in the storage and loss modulus, and a decrease in crystallinity and melt flow index with nanoclay content was also found. Although the presence of agglomerates can hinder the enhancement of Gd1 as it promotes agglomerate fracture and debonding, the increase in energy consumption through fibrillation and crazing promoted by the nanoclay prevails, suggesting that the nanoclay's toughening effect that has been extensively reported under quasi-static and impact tests, is also present under RCP conditions, and that the HDPE nanocomposites could be used in applications in which RCP must be prevented.

The pharmacokinetics and analgesic effects of extended-release buprenorphine administered subcutaneously in healthy dogs.

Buprenorphine is a partial µ agonist opioid used for analgesia in dogs. An extended-release formulation (ER-buprenorphine) has been shown to provide effective analgesia for 72 hr in rats and mice. Six healthy mongrel dogs were enrolled in a randomized, blinded crossover design to describe and compare the pharmacokinetics and pharmacodynamics of ER-buprenorphine administered subcutaneous at 0.2 mg/kg (ER-B) and commercially available buprenorphine for injection intravenously at 0.02 mg/kg (IV-B). After drug administration, serial blood samples were collected to measure plasma buprenorphine concentrations using liquid chromatography/mass spectrometry detection. Heart rate, respiratory rate, body temperature, sedation score, and thermal threshold latency were recorded throughout the study. Median (range) terminal half-life, time to maximum concentration, and maximum plasma concentration of ER-buprenorphine were 12.74 hr (10.43-18.84 hr), 8 hr (4-36 hr), and 5.00 ng/ml (4.29-10.98 ng/ml), respectively. Mild bradycardia, hypothermia, and inappetence were noted in both groups. Thermal threshold latency was significantly prolonged compared to baseline up to 12 hr and up to 72 hr in IV-B and ER-B, respectively. These results showed that ER-buprenorphine administered at a dose of 0.2 mg/kg resulted in prolonged and sustained plasma concentrations and antinociceptive effects up to 72 hr after drug administration.

From matrix nano- and micro-phase tougheners to composite macro-properties.

In this paper, firstly, the morphology and toughness of a range of bulk epoxy polymers, which incorporate a second phase of well-dispersed silica nanoparticles and/or rubber microparticles, have been determined. Secondly, the macro-properties of natural-fibre reinforced-plastic (NFRP) composites based upon these epoxy polymers have been ascertained, using (i) unidirectional flax fibres or (ii) regenerated-cellulose fibres in the architecture of a plain-woven fabric. Thirdly, the toughening mechanisms which are induced in these materials by the presence of the silica nanoparticles, the rubber microparticles and the natural fibres have been identified. Finally, the values of the toughness of the bulk epoxy polymers and corresponding NFRPs have been quantitatively modelled. The increased toughness recorded for the bulk epoxy polymer due to the presence of the silica nanoparticles and/or rubber microparticles was indeed typically transferred to the NFRP composites when using such epoxies as the matrices for the fibres. Thus, the important role that may be played by modifications to the epoxy matrices in order to increase the toughness of the composites was very clearly demonstrated by these results. However, notwithstanding, the toughening mechanisms induced by the fibres were essentially responsible for the very high toughnesses of the NFRP composites, compared with the bulk epoxy polymers. The modelling studies successfully predicted the values of toughness of the bulk epoxy polymers and of the NFRP composites. These studies also quantified the extent to which each toughening mechanism, induced by the second-phase nano- and microparticles and the natural fibres, contributed to the overall values of toughness of the materials. This article is part of the themed issue 'Multiscale modelling of the structural integrity of composite materials'.

A fatigue assessment technique for modular and pre-stressed orthopaedic implants.

Orthopaedic implants experience large cyclic loads, and pre-clinical analysis is conducted to ensure they can withstand millions of loading cycles. Acetabular cup developments aim to reduce wall thickness to conserve bone, and this produces high pre-stress in modular implants. As part of an implant development process, we propose a technique for preclinical fatigue strength assessment of modular implants which accounts for this mean stress, stress concentrating features and material processing. A modular cup's stress distributions were predicted computationally, under assembly and in vivo loads, and its cyclic residual stress and stress amplitude were calculated. For verification against damage initiation in low-cycle-fatigue (LCF), the peak stress was compared to the material's yield strength. For verification against failure in high-cycle-fatigue (HCF) each element's reserve factor was calculated using the conservative Soderberg infinite life criterion. Results demonstrated the importance of accounting for mean stress. The cup was predicted to experience high cyclic mean stress with low magnitude stress amplitude: a low cyclic load ratio (Rl=0.1) produced a high cyclic stress ratio (Rs=0.80). Furthermore the locations of highest cyclic mean stress and stress amplitude did not coincide. The minimum predicted reserve factor Nf was 1.96 (HCF) and 2.08 (LCF). If mean stress were neglected or if the stress ratio were assumed to equal the load ratio, the reserve factor would be considerably lower, potentially leading to over-engineering, reducing bone conservation. Fatigue strength evaluation is only one step in a broader development process, which should involve a series of verifications with the full range of normal and traumatic physiological loading scenarios, with representative boundary conditions and a representative environment. This study presents and justifies a fatigue analysis methodology which could be applied in early stage development to a variety of modular and pre-stressed prosthesis concepts, and is particularly relevant as implant development aims to maximise modularity and bone conservation.

Added value of narrow band imaging and confocal laser endomicroscopy in detecting Barrett's esophagus neoplasia.

BACKGROUND AND STUDY AIMS: Advances in endoscopic imaging techniques have enabled more accurate identification of subtle mucosal abnormalities. The aim of the study was to assess the accuracy of predicting high grade dysplasia (HGD) and intramucosal cancer (IMC) in mucosa predicted as being nondysplastic vs. dysplastic by high definition white light endoscopy (HD-WLE), narrow band imaging (NBI), and confocal laser endomicroscopy (CLE). PATIENTS AND METHODS: A cross-sectional study was performed in a tertiary referral setting between February 2010 and September 2011. A total of 50 consecutive patients who were referred to St Vincent's Hospital for management of dysplastic Barrett's esophagus were included. A prediction of likely histology was made for each mucosal point (four-quadrant every 1 cm and any visible mucosal abnormality), first with HD-WLE, followed by NBI, and finally CLE. Biopsies were taken at all of these points. RESULTS: A total of 1190 individual biopsy points were assessed. At histology, 39 biopsy points were found to harbor HGD and 52 biopsy points harbored IMC. For the detection of HGD/IMC the sensitivity, specificity, and accuracy were: HD - WLE, 79.1 %, 83.1 %, and 82.8 %; NBI, 89.0 %, 80.1 %, and 81.4 %; and CLE, 75.7 %, 80.0 %, and 79.9 %, respectively. All mucosal points with IMC and all patients with HGD were detected by targeted biopsies guided by HD-WLE and NBI without the need for random Seattle protocol biopsies. CONCLUSIONS: HD-WLE in combination with NBI is highly accurate in the detection of HGD/IMC. Performing targeted biopsies in the surveillance of Barrett's esophagus is possible in expert centers.

The influence of acetabular cup material on pelvis cortex surface strains, measured using digital image correlation.

Acetabular cup loosening is a late failure mode of total hip replacements, and peri-prosthetic bone deterioration may promote earlier failure. Preservation of supporting bone quality is a goal for implant design and materials selection, to avoid stress shielding and bone resorption. Advanced polymer composite materials have closer stiffness to bone than metals, ceramics or polymers, and have been hypothesised to promote less adverse bone adaptation. Computer simulations have supported this hypothesis, and the present study aimed to verify this experimentally. A composite hemi-pelvis was implanted with Cobalt Chromium (CoCr), polyethylene (UHMWPE) and MOTIS(®)carbon-fibre-reinforced polyether etherketone (CFR-PEEK) acetabular cups. In each case, load was applied to the implanted pelvis and Digital Image Correlation (DIC) was used for surface strain measurement. The test was repeated for an intact hemi-pelvis. Trends in implanted vs. intact bone principal strains were inspected to assess the average principal strain magnitude change, allowing comparison of the potential bone responses to implantation with the three cups. The CFR-PEEK cup was observed to produce the closest bone strain to the intact hip in the main load path, the superior peri-acetabular cortex (+12% on average, R(2)=0.84), in comparison to CoCr (+40%, R(2)=0.91) and UHWMPE cups (-26%, R(2)=0.94). Clinical observations have indicated that increased periacetabular cortex loading may result in reduced polar cancellous bone loading, leading to longer term losses in periprosthetic bone mineral density. This study provides experimental evidence to verify previous computational studies, indicating that cups produced using materials with stiffness closer to cortical bone recreate physiological cortical bone strains more closely and could, therefore, potentially promote less adverse bone adaptation than stiffer press-fitted implants in current use.

Pre-clinical evaluation of ceramic femoral head resurfacing prostheses using computational models and mechanical testing.

Ceramic-on-ceramic hip resurfacing can potentially offer the bone-conserving advantages of resurfacing while eliminating metal ion release. Thin-walled ceramic resurfacing heads are conceivable following developments in the strength and reliability of ceramic materials, but verification of new designs is required. The present study aimed to develop a mechanical pre-clinical analysis verification process for ceramic resurfacing heads, using the DeltaSurf prosthesis design as a case study. Finite element analysis of a range of in vivo scenarios was used to design a series of physiologically representative mechanical tests, which were conducted to verify the strength of the prosthesis. Tests were designed to simulate ideal and worst-case in vivo loading and support, or to allow comparison with a clinically successful metallic device. In tests simulating ideal loading and support, the prosthesis sustained a minimum load of 39 kN before fracture, and survived 10 000 000 fatigue cycles of 0.534 kN to 5.34 kN. In worst-case tests representing a complete lack of superior femoral head bone support or pure cantilever loading of the prosthesis stem, the design demonstrated strength comparable to that of the equivalent metal device. The developed mechanical verification test programme represents an improvement in the state of the art where international test standards refer largely to total hip replacement prostheses. The case study's novel prosthesis design performed with considerable safety margins compared with extreme in vivo loads, providing evidence that the proposed ceramic resurfacing heads should have sufficient strength to perform safely in vivo. Similar verification tests should be designed and conducted for novel ceramic prosthesis designs in the future, leading the way to clinical evaluation.

Variable stocking effect and endemic population genetic structure in Murray cod Maccullochella peelii.

Microsatellite markers were utilized to examine the genetic structure of Murray cod Maccullochella peelii throughout its distribution in the Murray--Darling Basin (MDB) of eastern Australia, and to assess the genetic effects of over three decades of stocking hatchery-reared fingerlings. Bayesian analysis using the programme Structure indicated that the species is largely genetically panmictic throughout much of its extensive range, most probably due to the high level of connectivity between catchments. Three catchments with terminal wetlands (the Lachlan, Macquarie and Gwydir), however, contained genetically distinct populations. No stocking effects were detected in the catchments that were genetically panmictic (either because of low genetic power or lack of effects), but the genetically differentiated Gwydir and Macquarie catchment populations were clearly affected by stocking. Conversely, there was no genetic evidence for survival and reproduction of stocked fish in the Lachlan catchment. Therefore, stocking of M. peelii throughout the MDB has resulted in a range of genetic effects ranging from minimal detectable effect, to substantial change in wild population genetic structure.

Biopsy forceps is inadequate for the resection of diminutive polyps.

BACKGROUND AND STUDY AIMS: Cold biopsy forceps polypectomy (CBP) is often used for the removal of diminutive polyps. The efficacy of the technique has not been thoroughly assessed. The aim of this study was to prospectively assess the efficacy of CBP for removing diminutive polyps. PATIENTS AND METHODS: This was a prospective study from St Vincent's Hospital, a tertiary referral hospital in Melbourne, Australia. A total of 143 patients were screened and 52 patients with ≥ 1 diminutive polyps were enrolled. CBP was used to resect diminutive polyps until no polyp tissue was visible. The polyp base was then resected using endoscopic mucosal resection (EMR) with a 1 - 2-mm margin. The CBP and EMR samples were compared to assess completeness of the resection. RESULTS: Overall 39 % (21 / 54) of diminutive polyps were completely resected using CBP. After binary logistic regression analysis, polyp histology was found to be predictive of resection, with complete resection of 62 % (13 / 21) for adenomas and 24 % (8 / 33) for hyperplastic polyps (odds ratio 5.1; P = 0.008). The size and number of bites taken with the forceps were not predictive of complete response. CONCLUSIONS: Within the limitations of a modest sample size, CBP appears to be inadequate treatment for the removal of diminutive polyps.

Experimental validation of a finite element model of the proximal femur using digital image correlation and a composite bone model.

Computational biomechanical models are useful tools for supporting orthopedic implant design and surgical decision making, but because they are a simplification of the clinical scenario they must be carefully validated to ensure that they are still representative. The goal of this study was to assess the validity of the generation process of a structural finite element model of the proximal femur employing the digital image correlation (DIC) strain measurement technique. A finite element analysis model of the proximal femur subjected to gait loading was generated from a CT scan of an analog composite femur, and its predicted mechanical behavior was compared with an experimental model. Whereas previous studies have employed strain gauging to obtain discreet point data for validation, in this study DIC was used for full field quantified comparison of the predicted and experimentally measured strains. The strain predicted by the computational model was in good agreement with experimental measurements, with R(2) correlation values from 0.83 to 0.92 between the simulation and the tests. The sensitivity and repeatability of the strain measurements were comparable to or better than values reported in the literature for other DIC tests on tissue specimens. The experimental-model correlation was in the same range as values obtained from strain gauging, but the DIC technique produced more detailed, full field data and is potentially easier to use. As such, the findings supported the validity of the model generation process, giving greater confidence in the model's predictions, and digital image correlation was demonstrated as a useful tool for the validation of biomechanical models.

Performance of the resurfaced hip. Part 2: The influence of prosthesis stem design on remodelling and fracture of the femoral neck.

Hip resurfacing is a popular treatment for osteoarthritis in young, active patients. Previous studies have shown that occasional failures--femoral neck fracture and implant loosening, possibly associated with bone adaptation--are affected by prosthesis sizing and positioning, in addition to patient and surgical factors. With the aim of improving tolerance to surgical variation, finite element modelling was used to indicate the effects of prosthesis metaphyseal stem design on bone remodelling and femoral neck fracture, with a range of implant orientations. The analysis suggested that the intact femoral neck strength in trauma could be maintained across a wider range of varus-valgus orientations for short-stemmed and stemless prostheses. Furthermore, the extent of periprosthetic bone remodelling was lower for the short-stemmed implant, with slightly reduced stress shielding and considerably reduced densification around the stem, potentially preventing further progressive proximal stress shielding. The study suggests that a short-stemmed resurfacing head offers improved tolerance to misalignment and remodelling stimulus over traditional designs. While femoral neck fracture and implant loosening are multifactorial, biomechanical factors are of clear importance to the clinical outcome, so this may reduce the risk for patients at the edge of the indications for hip resurfacing, or shorten the surgical learning curve.

Building leadership capacity to drive sustainable water management: the evaluation of a customised program.

This paper describes a customised, six-month, leadership development program (LDP) that was designed for emerging leaders in the Australian water industry who were promoting sustainable urban water management (SUWM). It also presents results from an evaluation of the program's benefits, costs and overall 'return on investment' (ROI). The program was designed to help build emergent leadership capacity in the water industry, given strong evidence that this form of leadership plays an important role in advancing SUWM. It involved '360-degree feedback' processes, training, individual leadership development plans, and coaching sessions. Its design was informed by a review of the literature, and its content was informed by local empirical research involving effective SUWM leaders. The evaluation used a seven-tier assessment framework that examined different dimensions of the program's performance using source and methodological triangulation. The results indicate that such LDPs can produce a range of positive outcomes, such as promoting desired leadership behaviours and generating a positive ROI estimate. Specifically, the program's estimated ROI was approximately 190% after only one year. The primary conclusion is that evidence-based LDPs which are highly customised for specific types of leaders in the water industry represent a promising type of intervention to build forms of leadership capacity which are needed to successfully promote SUWM.

Performance of the resurfaced hip. Part 1: the influence of the prosthesis size and positioning on the remodelling and fracture of the femoral neck.

Hip resurfacing is an established treatment for osteoarthritis in young active patients. Failure modes include femoral neck fracture and prosthesis loosening, which may be associated with medium-term bone adaptation, including femoral neck narrowing and densification around the prosthesis stem. Finite element modelling was used to indicate the effects of prosthesis sizing and positioning on the bone remodelling and fracture strength under a range of normal and traumatic loads, with the aim of understanding these failure modes better. The simulations predicted increased superior femoral neck stress shielding in young patients with small prostheses, which required shortening of the femoral neck to give an acceptable implant-bone interface. However, with a larger prosthesis, natural femoral head centre recreation in the implanted state was possible; therefore stress shielding was restricted to the prosthesis interior, and its extent was less sensitive to prosthesis orientation. With valgus orientation, the implanted neck strength was, at worst, within 3 per cent of its intact strength. The study suggests that femoral neck narrowing may be linked to a reduction in the horizontal femoral offset, occurring if the prosthesis is excessively undersized. As such, hip resurfacing should aim to reproduce the natural femoral head centre, and, for valgus prosthesis orientation, to avoid femoral neck fracture.

Characterisation of dihydrodipicolinate synthase (DHDPS) from Bacillus anthracis.

Bacillus anthracis is a Gram-positive spore-forming bacterium that is the causative agent of anthrax disease. The use of anthrax as a bioweapon has increased pressure for the development of an effective treatment. Dihydrodipicolinate synthase (DHDPS) catalyses the first committed step in the biosynthetic pathway yielding two essential bacterial metabolites, meso-diaminopimelate (DAP) and (S)-lysine. DHDPS is therefore a potential antibiotic target, as microbes require either lysine or DAP as a component of the cell wall. This paper is the first biochemical description of DHDPS from B. anthracis. Enzyme kinetic analyses, isothermal titration calorimetry (ITC), mass spectrometry and differential scanning fluorimetry (DSF) were used to characterise B. anthracis DHDPS and compare it with the well characterised Escherichia coli enzyme. B. anthracis DHDPS exhibited different kinetic behaviour compared with E. coli DHDPS, in particular, substrate inhibition by (S)-aspartate semi-aldehyde was observed for the B. anthracis enzyme (K(si(ASA))=5.4+/-0.5 mM), but not for the E. coli enzyme. As predicted from a comparison of the X-ray crystal structures, the B. anthracis enzyme was not inhibited by lysine. The B. anthracis enzyme was thermally stabilised by the first substrate, pyruvate, to a greater extent than its E. coli counterpart, but has a weaker affinity for pyruvate based on enzyme kinetics and ITC studies. This characterisation will provide useful information for the design of inhibitors as new antibiotics targeting B. anthracis.

Sustainable urban water management: understanding and fostering champions of change.

This paper highlights and discusses ten characteristic attributes of emergent leaders (also known as 'champions') who worked as influential change agents within publicly managed, Australian water agencies to encourage more sustainable forms of urban water management. These attributes relate to: the 'openness to experience' personality characteristic; career mobility and work history demographics; personal and position power; strategic social networks; the culture of their organisations; and five distinguishing leadership behaviours (e.g. persisting under adversity). Guided by the findings of an international literature review, the author conducted a multiple case study involving six water agencies. This research identified attributes of these leaders that were typically strong and/or distinguishing compared to relevant control groups, as well as influential contextual factors. While it is widely acknowledged that these leaders play a critical role in the delivery of sustainable urban water management, there has been a paucity of context-sensitive research involving them. The research project highlighted in this paper is a response to this situation and has led to the development of a suite of 39 practical, evidence-based strategies to build leadership capacity throughout water agencies. Such capacity is one of the elements needed to drive the transition to more 'water sensitive cities'.

Leader of the pack: faecal pellet deposition order impacts PCR amplification in wombats.

DNA sourced from faeces is notoriously less reliable than that from tissue. Hence, understanding whether faecal pellet quality varies within faecal piles may be important for sample selection. We hypothesized that the order in which faecal pellets are deposited may influence microsatellite polymerase chain reaction (PCR) amplification success from sampled faeces, more specifically, that first pellets deposited will have signatures of greater success than later ones. In a first test of the hypothesis, first and later-deposited pellets, as determined from the direction of footprints, were collected from fresh (overnight) faecal piles of northern hairy-nosed wombats (Lasiorhinus krefftii). DNA extracts were typed for seven microsatellite loci. We found that faecal deposition order significantly affected optical density of bands on autoradiographs (a measure of PCR amplification success) when the first faecal pellet was compared with the last one, but not when the first pellet was only distinguishable from later ones. The absence of a difference in amplification rate between first and later pellets is likely a reflection of the overall high amplification success in this study. That first pellets deposited yield more product suggests they contain more intestinal cells. Although further comparisons are needed, these results may inform sample selection in species for which success of microsatellite PCR amplification of faecal DNA is low. Deposition order may have more of an impact on amplification success and genotyping errors as faecal age increases.

An Interesting Case of Recurrent Small Bowel Obstruction.

Sclerosing mesenteritis is associated with a spectrum of diseases which include mesenteric lipodystrophy and mesenteric panniculitis. This inflammatory and fibrosing disorder can affect the small and large bowel wall and mesenteric vessels by exerting a mass effect. The following case highlights the difficulties with diagnosing and managing this unusual disease. A 64-year-old man presented with acute central abdominal pain, radiating to his back, and profuse vomiting. He was diagnosed clinically with small bowel obstruction. He had had an episode of small bowel obstruction 6 years earlier. At this time, he underwent an exploratory laparotomy, and a mass was identified in the small bowel mesentery. The features were thought to be in keeping with sclerosing mesenteritis. He had a dramatically favourable response to the initiation of prednisolone. He continued to be well and asymptomatic for a further 5 years on long-term maintenance low-dose steroids and 6-mercaptopurine. He re-presented in 2009 (six years after initial presentation) with very severe acute abdominal pain and vomiting. He had no recent change in weight or appetite, and had not had time off work. He underwent a second laparotomy and the tissue diagnosis was of metastatic carcinoid tumour involving the small bowel mesentery. This is the first case to our knowledge where sclerosing mesenteritis has been confirmed histologically on biopsy and then subsequently diagnosed with histologically proven carcinoid tumour. For this particular reason it must be always remembered that sclerosing mesenteritis is a 'pathological' and not a radiological diagnosis and that a large proportion of cases are associated with neoplasia.

Extracellular proteases and possible disease related virulence mechanisms of two marine bacteria implicated in an opportunistic bacterial infection of Nephrops norvegicus.

The extracellular products (ECP) secreted by two strains of gram-negative bacteria isolated from Nephrops norvegicus exhibiting signs of an opportunistic bacterial infection were investigated with the objective of understanding their role in the spoilage of host muscle tissue and identifying disease related virulence mechanisms. ECP from Vibrio sp. demonstrated no proteolytic activity. ECP from Pseudoalteromonas sp. (isolate N10) degraded several substrates, including azocasein and host muscle tissue. Proteolytic activity increased with temperature. Substrate-impregnated sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the effect of the isolates' ECP on the molecular weight of proteins derived from abdominal muscle tissue revealed that the ECP of Pseudoalteromonas sp. selectively degraded the myosin heavy chain, troponin-T, troponin-I, paramyosin and several unidentified muscle proteins approximately 110 kDa in size. Topomyosin was also reduced in quantity. Degradation of SDS-PAGE gels impregnated with host muscle proteins, by the ECP of Pseudoalteromonas sp. revealed 3 zones of proteolysis, with estimated molecular weights between 100 and 30 kDa, indicating multiple proteases in the ECP. Through the API ZYM system, both isolates demonstrated strong leucine arylamidase activity, with the Vibrio sp. showing strong acid phosphatase activity. These enzymes have been identified as disease related virulence mechanisms in other bacterial pathogens. There is likely a complex pathway to the final condition, involving virulence factors of other species and the stresses involved in capture and transport.

Idiopathic muscle necrosis in the Norway lobster, Nephrops norvegicus (L.): aetiology, pathology and progression to bacteraemia.

The pathology and progression of idiopathic muscle necrosis (IMN) in Nephrops norvegicus and possible aetiologies have been investigated. Trawl capture, aerial exposure and handling initiate IMN, and the condition can be induced through periods of aerial exposure alone, in the absence of trawling. Within 24-48 h after trawl capture IMN progresses to a multi-species bacterial septicaemia, with moribund animals exhibiting clinical signs. The aetiology of this condition has been examined using molecular (16S rRNA gene sequencing) and biochemical (standard taxonomic assays, Biolog) criteria to characterize bacterial isolates from moribund and healthy animals. Histopathology of the IMN phase reveals a loss of sarcomeric structure with necrotic lesions containing pyknotic nuclei, fragments of myofibrils and connective tissue elements. In the bacterial phase there is extensive loss of abdominal muscle structure, and the presence of rod-shaped Gram-negative bacteria in the degrading tissues. The results demonstrate that the IMN condition is connected to stressful conditions imposed on N. norvegicus, but involves no pathogenic agents. This is followed by an opportunistic bacterial infection that causes further tissue spoilage. It is believed that the primary cause of both IMN and bacteraemia is imposed stress, but they are expressed in different time courses.

Phylogeography and environmental correlates of a cap on reproduction: teat number in a small marsupial, Antechinus agilis.

Natural selection should optimize litter size in response to the distribution and abundance of resources during breeding. In semelparous, litter-bearing antechinuses, teat number limits litter size. Consequently adaptation has been inferred in explaining intraspecific, geographic variability in teat number for several Antechinus spp. The phylogeography of teat number variation and associated genetic divergence were assessed in A. agilis using nine microsatellites and mitochondrial cytochrome b sequence data. Six-teat Otway Range animals were divergent in microsatellite allele identity and frequencies: samples from three Otway six-teat sites demonstrated significantly greater similarity genetically to those from six-teat animals approximately 250 km to the west, than to nearby Otway 10-teat samples, or to the six-teat animals at Wilsons Promontory. Gene flow between Otway phenotypes appears to have been limited for sufficient time to enable different microsatellite alleles to evolve. Nonetheless, nuclear genetic evidence suggested only incomplete reproductive isolation, and mitochondrial DNA (mtDNA) haplotypes showed no association with teat number. Other populations across the range were no more genetically differentiated from one another than expected from geographic separation. Principal components and distance-based redundancy analyses found an association between environmental variables and geographic distribution of A. agilis teat number - six-teat animals inhabit more temperate forests, whilst those with more teats experience greater seasonality. The apparent restricted breeding between phenotypically distinct animals, together with phylogenetically separate groups of six-teat animals in different locations with similar environments, are consistent with the hypothesis that adaptation to different habitats drives teat number variation in A. agilis.