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Estuarine ecosystems are threatened by numerous anthropogenic pressures. Fish assemblages are a dominant component of estuarine macrofauna and serve as indicators for the health of these transitional water ecosystems. Environmental DNA (eDNA) metabarcoding is increasingly used to assess the biodiversity of fishes in estuaries. However, there is a need to further establish how effective eDNA metabarcoding can be relative to conventional fish sampling methods across multiple estuaries and seasons. This study compared fish assemblages detected via eDNA metabarcoding of surface water samples to contemporary sampling with conventional fishing gears in three temperate estuaries (UK), during early summer and autumn. Most species caught by fishing were detected by eDNA. Species richness estimates from eDNA were two to ten times higher than estimates based on fishing, and included taxa of conservation importance and a non-native species. The eDNA assemblage composition was significantly different to the assemblage detected by seine nets. Importantly, eDNA methods could effectively discriminate between fish assemblages of different estuaries and seasons. Fish assemblages in estuaries are often not monitored due to resource constraints. The dynamic nature of estuaries may make fishing gear deployment difficult and inconsistent. The findings indicate that eDNA metabarcoding is suited to gathering large amounts of information on fish biodiversity, at a relatively low sampling effort, compared to established fishing methods. Therefore, eDNA shows promise as an assessment tool for fish assemblage structure and ecosystem health in estuarine environments, with application to statutory monitoring.

Recurrent reproductive failure (RRF) encompasses recurrent implantation failure (RIF) and recurrent pregnancy loss (RPL). These highly prevalent, distressing conditions have many unanswered questions regarding aetiology and management. Oestrogen receptor beta (ERβ) is the predominant oestrogen receptor expressed in the vascular endothelium of the endometrium during the window of implantation (WOI). The establishment of normal endometrial receptivity is integrally associated with progesterone receptor (PR). Therefore, we aimed to investigate whether women with RRF have clinical, type-specific endometrial aberrations of ERβ, PR and Ki-67 expression during the WOI. Thirty-eight endometrial biopsies were collected; 29 RRF (10 RIF, 9 recurrent loss of early pregnancy (RLEP) and 10 recurrent fetal loss (RFL)) and 9 fertile controls (FC). Within RIF, RLEP and RFL groups, the perivascular compartment showed significantly lower levels of ERβ vs FC (P = 0.02, P = 0.03 and P = 0.01, respectively). Vascular endothelium also displayed significantly lower levels of ERβ within RIF and RFL cohorts vs FC (P = 0.03 and P = 0.003). The expression of Ki-67 was significantly lower within vascular endothelium of all RRF; RIF (P = 0.02), RLEP (P = 0.02) and RFL (P < 0.01). PR was significantly reduced (P < 0.001) in the perivascular area of women with RIF. These findings provide novel insights into biological correlates of clinical subtypes of RRF. The endometrium of women with RRF display significantly altered levels of ERβ, PR and Ki-67 during the WOI, furthering our understanding of the defective endometrial phenotype of women suffering from RRF, with possible impaired glandular function, angiogenesis and decidualisation.

Aquaculture growth is hindered by an increasing number of challenges, primarily infectious diseases and inappropriate or unsustainable fish nutrition. Hence it is critical to develop novel prevention strategies to minimise infectious diseases and pharmaceutical interventions. Nutritional challenges and the health of the fish could be improved by managing their microbial communities. Microbiomes can play a crucial role in fish physiology, particularly in digestion, by metabolizing largely indigestible feed components for the host or synthesis essential micronutrients. Beyond their nutritional role, microbiomes are considered the first line of defence against pathogens. In this study, a novel prebiotic mix (Selectovit), composed of 1,3/1,6-beta glucans, mannan-oligosaccharides, nucleic acids, nucleotides, medium chain fatty acids and single chain fatty acids, was tested at different inclusion levels (0.0; 0.5; 1.0; 2.0 g/kg) in the diet of Atlantic salmon (Salmo salar). Using experimental feed trials and 16 S rRNA microbiome profiling, the impact of the prebiotic blend on fish growth and microbial community within both the gastrointestinal tract and the skin was assessed. Overall, the supplement showed no significant impact on growth. However, we clearly demonstrate that the prebiotic can significantly manipulate the microbial community of the distal intestine and the skin. Several potential beneficial bacteria such as Bacillus and Mycoplasma spp. were significantly more abundant in the prebiotic-fed groups compared to the control. In contrast, putative pathogenic bacteria were less abundant in the salmon fed the prebiotic blend. Interestingly, the supplement induced more changes in the skin than in the gut. There is growing evidence in fish for highly complex interactions between the microbial communities of the digestive system and external mucosa, and with the host immune system. Further research in this field could lead to the creation of novel bacterial biomarkers and new non-invasive strategies for fish digestive health monitoring.

Demand for underground railways has rapidly increased due to accelerated urbanisation and population growth. This has elevated the importance of tunnel designs with adequate fire safety and protection measures. However, due to intricate modern rail tunnel designs, prescriptive codes are often difficult to implement and lead to over-conservative design. In this study, the current state of tunnel fire analysis was reviewed with a focus on Australia. A large-eddy simulation (LES)-based fire model was applied to investigate the temperature and smoke dispersion from a 2 MW metro tunnel fire case scenario to the cross-passage. A total of 28 cases with various cross-passage ventilation settings were examined, including longitudinal tunnel velocity, cross-passage velocity, train location relative to the cross-passage and fire location. The modelling showed that a 0.84 m/s critical velocity was sufficient for smoke control in the cross-passage. Furthermore, two empirical methods for cross-passage critical velocity were performed, which showed utilisation of the Froude number produced a less conservative critical velocity (0.610 m/s) compared to the dimensionless method (0.734 m/s). Nevertheless, both numerical and empirical results were significantly lower than the standard 1.0 m/s minimum flow rate for smoke control (AS1668.1). The results provide preliminary evidence towards the need for revision of current tunnel fire standards and response protocols.

Elucidating the diets of insect predators is important in basic and applied ecology, such as for improving the effectiveness of conservation biological control measures to promote natural enemies of crop pests. Here, we investigated the aphid diet of two common aphid predators in Central European agroecosystems, the native Coccinella septempunctata (Linnaeus) and the invasive Harmonia axyridis (Pallas; Coleoptera: Coccinellidae) by means of high throughput sequencing (HTS). For acquiring insights into diets of mobile flying insects at landscape scale minimizing trapping bias is important, which imposes methodological challenges for HTS. We therefore assessed the suitability of three field sampling methods (sticky traps, pan traps and hand-collection) as well as new aphid primers for identifying aphid prey consumption by coccinellids through HTS. The new aphid primers facilitate identification to species level in 75% of the European aphid genera investigated. Aphid primer specificity was high in silico and in vitro but low in environmental samples with the methods used, although this could be improved in future studies. For insect trapping we conclude that sticky traps are a suitable method in terms of minimizing sampling bias, contamination risk and trapping success, but compromise on DNA-recovery rate. The aphid diets of both field-captured ladybird species were dominated by Microlophium carnosum, the common nettle aphid. Another common prey was Sitobion avenae (cereal aphid), which got more often detected in C. septempunctata compared to H. axyridis. Around one third of the recovered aphid taxa were common crop pests. We conclude that sampling methodologies need constant revision but that our improved aphid primers offer currently one of the best solutions for broad screenings of coccinellid predation on aphids.

Abstract Background For healthcare systems, an ageing population poses challenges in the delivery of equitable and effective care. Frailty assessment has the potential to improve care in the intensive care setting, but applying assessment tools in critical illness may be problematic. The aim of this systematic review was to evaluate evidence for the feasibility and reliability of frailty assessment in critical care. Methods Our primary search was conducted in Medline, Medline In-process, EMBASE, CINAHL, PsycINFO, AMED, Cochrane Database of Systematic Reviews, and Web of Science (January 2001 to October 2017). We included observational studies reporting data on feasibility and reliability of frailty assessment in the critical care setting in patients 16 years and older. Feasibility was assessed in terms of timing of evaluation, the background, training and expertise required for assessors, and reliance upon proxy input. Reliability was assessed in terms of inter-rater reliability. Results Data from 11 study publications are included, representing 8 study cohorts and 7761 patients. Proxy involvement in frailty assessment ranged from 58 to 100%. Feasibility data were not well-reported overall, but the exclusion rate due to lack of proxy availability ranged from 0 to 45%, the highest rate observed where family involvement was mandatory and the assessment tool relatively complex (frailty index, FI). Conventional elements of frailty phenotype (FP) assessment required modification prior to use in two studies. Clinical staff tended to use a simple judgement-based tool, the clinical frailty scale (CFS). Inter-rater reliability was reported in one study using the CFS and although a good level of agreement was observed between clinician assessments, this was a small and single-centre study. Conclusion Though of unproven reliability in the critically ill, CFS was the tool used most widely by critical care clinical staff. Conventional FP assessment required modification for general application in critical care, and an FI-based assessment may be difficult to deliver by the critical care team on a routine basis. There is a high reliance on proxies for frailty assessment, and the reliability of frailty assessment tools in critical care needs further evaluation. Prospero registration number CRD42016052073.

New infections of the amphibian chytrid fungus could arise from other animal reservoirs in the environment. Here, Liewet al. demonstrate that zebrafish can be infected by chytrid similarly to amphibians, expanding our understanding of how this pathogen can parasitize its hosts.