Effectiveness of antifungal treatments during chytridiomycosis epizootics in populations of an endangered frog

The recently-emerged amphibian chytrid fungus Batrachochytrium dendrobatidis (Bd) has had an unprecedented impact on global amphibian populations, and highlights the urgent need to develop effective mitigation strategies. We conducted in-situ antifungal treatment experiments in wild populations of the endangered mountain yellow-legged frog during or immediately after Bd-caused mass die-off events. The objective of treatments was to reduce Bd infection intensity (“load”) and in doing so alter frog-Bd dynamics and increase the probability of frog population persistence despite ongoing Bd infection. Experiments included treatment of early life stages (tadpoles and subadults) with the antifungal drug itraconazole, treatment of adults with itraconazole, and augmentation of the skin microbiome of subadults with Janthinobacterium lividum, a commensal bacterium with antifungal properties. All itraconazole treatments caused immediate reductions in Bd load, and produced longer-term effects that differed between life stages. In experiments focused on early life stages, Bd load was reduced in the 2 months immediately following treatment and was associated with increased survival of subadults. However, Bd load and frog survival returned to pre-treatment levels in less than 1 year, and treatment had no effect on population persistence. In adults, treatment reduced Bd load and increased frog survival over the entire 3-year post-treatment period, consistent with frogs having developed an effective adaptive immune response against Bd. Despite this protracted period of reduced impacts of Bd on adults, recruitment into the adult population was limited and the population eventually declined to near-extirpation. In the microbiome augmentation experiment, exposure of subadults to a solution of J. lividum increased concentrations of this potentially protective bacterium on frogs. However, concentrations declined to baseline levels within 1 month and did not have a protective effect against Bd infection. Collectively, these results indicate that our mitigation efforts were ineffective in causing long-term changes in frog-Bd dynamics and increasing population persistence, due largely to the inability of early life stages to mount an effective immune response against Bd. This results in repeated recruitment failure and a low probability of population persistence in the face of ongoing Bd infection.

Exploring reasons for recruitment failure in clinical trials: a qualitative study with clinical trial stakeholders in Switzerland, Germany, and Canada

Background Poor participant recruitment is the most frequent reason for premature discontinuation of randomized clinical trials (RCTs), particularly if they are investigator-initiated. The aims of this qualitative study were to investigate (1) the views of clinical trial stakeholders from three different countries regarding reasons for recruitment failure in RCTs and (2) how these compare and contrast with the causes identified in a previous systematic review of RCT publications. Methods From August 2015 to November 2016, we conducted 49 semi-structured interviews with a purposive sample of clinical trial stakeholders. This included investigators based in Germany (n = 9), Switzerland (n = 6) and Canada (n = 1) with personal experience of a discontinued RCT and 33 other stakeholders (e.g., representatives of ethics committees, clinical trial units, pharmaceutical industry) in Switzerland. Individual semi-structured qualitative interviews were conducted and analyzed using thematic analysis. Results Interviewees identified a total of 29 different reasons for recruitment failure. Overoptimistic recruitment estimates, too narrow eligibility criteria, lack of engagement of recruiters/trial team, lack of competence/training/experience of recruiters, insufficient initial funding, and high burden for trial participants were mentioned most frequently. The interview findings largely confirm the previous systematic review on published reasons for recruitment failure. However, eight new reasons for recruitment failure were identified in the interviews, which led to the checklist of reasons for recruitment failure being revised and a new category describing research environment-related factors being added. Conclusions This study highlights the diversity of often interlinked reasons for recruitment failure in RCTs. Integrating the findings of this interview study with a previous systematic review of RCT publications led to a comprehensive, structured checklist of empirically-informed reasons for recruitment failure. The checklist may be useful to guide further research on interventions to improve participant recruitment in RCTs and helpful for trial investigators, research ethics committees, and funding agencies when assessing trial feasibility with respect to recruitment.

Climate change, fire return intervals and the growing risk of permanent forest loss in boreal Eurasia

Climate change has driven an increase in the frequency and severity of fires in Eurasian boreal forests. A growing number of field studies have linked the change in fire regime to post-fire recruitment failure and permanent forest loss. In this study we used four burnt area and two forest loss datasets to calculate the landscape-scale fire return interval (FRI) and associated risk of permanent forest loss. We then used machine learning to predict how the FRI will change under a high emissions scenario (SSP3-7.0) by the end of the century. We found that there is currently 133 000 km2 at high, or extreme, risk of fire-induced forest loss, with a further 3 M km2 at risk by the end of the century. This has the potential to degrade or destroy some of the largest remaining intact forests in the world, negatively impact the health and economic wellbeing of people living in the region, as well as accelerate global climate change.

Barriers to recruitment to an orthopaedic randomized controlled trial comparing two surgical procedures for ankle arthritis

Aims A multicentre, randomized, clinician-led, pragmatic, parallel-group orthopaedic trial of two surgical procedures was set up to obtain high-quality evidence of effectiveness. However, the trial faced recruitment challenges and struggled to maintain recruitment rates over 30%, although this is not unusual for surgical trials. We conducted a qualitative study with the aim of gathering information about recruitment practices to identify barriers to patient consent and participation to an orthopaedic trial. Methods We collected 11 audio recordings of recruitment appointments and interviews of research team members (principal investigators and research nurses) from five hospitals involved in recruitment to an orthopaedic trial. We analyzed the qualitative data sets thematically with the aim of identifying aspects of informed consent and information provision that was either unclear, disrupted, or hindered trial recruitment. Results Recruiters faced four common obstacles when recruiting to a surgical orthopaedic trial: patient preferences for an intervention; a complex recruitment pathway; various logistical issues; and conflicting views on equipoise. Clinicians expressed concerns that the trial may not show significant differences in the treatments, validating their equipoise. However, they experienced role conflicts due to their own preference and perceived patient preference for an intervention arm. Conclusion This study provided initial information about barriers to recruitment to an orthopaedic randomized controlled trial. We shared these findings in an all-site investigators’ meeting and encouraged researchers to find solutions to identified barriers; this led to the successful completion of recruitment. Complex trials may benefit for using of a mixed-methods approach to mitigate against recruitment failure, and to improve patient participation and informed consent. Cite this article: Bone Jt Open 2021;2(8):631–637.

Explaining Recruitment Stochasticity at a Species’ Range Margin

Advancing our understanding of how environmental variability affects the distribution of organisms is crucial for ecology and conservation. The exploration of changes in demographic patterns close to species distribution margins is important as populations here may provide a window into future population changes also elsewhere. However, the knowledge of factors causing recruitment variation is still inadequate in many systems and this deficiency is particularly evident close to species’ distribution borders. We studied the spatiotemporal variability in recruit-adult dynamics in a blue mussel, Mytilus trossulus, population to get insights into how environmental variables drive variation in recruitment and how this variability affects adult population growth. Thirty sites along a wave exposure gradient were monitored during four consecutive years. From each site, mussels were collected both from artificial recruitment units and from natural mussel beds. Our results showed high year-to-year variation in recruitment strength with high spatial variation. Mussel recruitment to artificial units and later recruitment to the benthos correlated highly. Juvenile abundances 1 year later paralleled prior recruitment strengths and caused synchronous but time-lagged changes in adult cohorts. Seawater salinity was the strongest predictor for recruitment variation, whereas sea temperature and wave exposure had low predictive power for this early life stage. For juveniles and for adults in the benthos, wave exposure explained the variation best, whereas temperature and especially salinity explained less. The results indicate that (a) the studied blue mussel population is strongly driven by variation in recruitment strength that (b) drives the size of the later cohorts, and the population is possibly even (c) recruitment limited in some years. Our study predicts a challenging future for this range population, resulting from a higher frequency of recruitment failure caused by a deteriorating sea climate. Knowledge about factors underlying variation in recruitment is thus essential for forecasting the future of this range population and for conserving its future state.

Changes in Panulirus cygnus Settlement Along Western Australia Using a Long Time Series

The pelagic development stages of many marine invertebrate species dictates their spatial and temporal distribution once reaching their benthic second phase of life. This life cycle is associated with the Western Rock Lobster (Panulirus cygnus) along the coast of Western Australia. Over the past 50 years, the number of puerulus reaching the nearshore reefs after their first 9 to 11 months of pelagic life in Western Australia has been monitored. These numbers, collected now at eight sites over the latitudes of the fishery, are indicative of the catchable stock 3–4 years into the future. In 2008, the fishery experienced a recruitment failure which lasted for several years before recovering to mean numbers pre-2008. This was associated with spatial and temporal shifts in the patterns of puerulus settlement. Previous research has hypothesized that physical and biological conditions in the south-east Indian Ocean no longer favored their survival. However, this decline has not been attributed to a single process. As the recovery is ongoing, contrasts in the settlement data before and after the decline are not completed. Here we characterize the data using ANOVA and pairwise comparisons to gain a better understanding of the typical patterns after the decline. Our results demonstrate that there has been a significant reduction in puerulus numbers over the first half of the season at all sites post decline. For the sites south of Lancelin there has been a significant reduction in puerulus numbers over the whole season. In addition, sites that show signs of recovery indicate that the majority of settlement occurred in the second half of the season. We anticipate these results to be the starting point for focused research into the environmental changes which may have occurred to generate these shifts in settlement numbers both from a timing and spatial perspective.

Effectiveness of antifungal treatments during chytridiomycosis epizootics in populations of an endangered frog

The recently-emerged amphibian chytrid fungus Batrachochytrium dendrobatidis (Bd) has had an unprecedented impact on global amphibian populations, and highlights the urgent need to develop effective mitigation strategies against this pathogen. We conducted field antifungal treatment experiments in populations of the endangered mountain yellow-legged frog during or immediately after Bd-caused mass die-off events. The objective of the treatments was to reduce Bd infection intensity ("load") and in doing so alter frog-Bd dynamics and increase the probability of frog population persistence despite ongoing Bd infection. Experiments included treatment of early life stages (tadpoles and subadults) with the antifungal drug itraconazole, treatment of adults with itraconazole, and augmentation of the skin microbiome of subadults with Janthinobacterium lividum, a commensal bacterium with antifungal properties. All itraconazole treatments caused immediate reductions in Bd load, and produced longer-term effects that differed between life stages. In experiments focused on early life stages, Bd load was reduced in the two months immediately following treatment and was associated with increased survival of subadults. However, Bd load and frog survival returned to pre-treatment levels in less than one year, and treatment had no effect on population persistence. In adults, treatment reduced Bd load and increased frog survival over the three-year post-treatment period, consistent with frogs having developed an effective adaptive immune response against Bd. Despite this protracted period of reduced impacts of Bd on adults, recruitment of new individuals into the adult population was limited and the population eventually declined to near-extirpation. In the microbiome augmentation experiment, bathing frogs in a J. lividum solution after Bd load reduction with itraconazole increased concentrations of this bacterium on frogs, but concentrations declined to baseline levels within one month and did not have a protective effect against Bd infection. Collectively, these results suggest that Bd mitigation efforts focused on frog populations that have recently declined due to Bd emergence are ineffective in causing long-term changes in frog-Bd dynamics and increasing population persistence, due largely to the inability of early life stages to mount an effective immune response against Bd and resulting high susceptibility. This results in repeated recruitment failure and a low probability of population persistence in the face of ongoing Bd infection.

Using an oceanographic model to investigate the mystery of the missing puerulus

Dynamics of ocean boundary currents and associated shelf processes can influence onshore/offshore transport of water, critically impacting marine organisms that release long-lived pelagic larvae into the water column. The western rock lobster, Panulirus cygnus, endemic to Western Australia, is the basis of Australia's most valuable wild-caught commercial fishery. After hatching, western rock lobster larvae (phyllosoma) spend up to 11 months in offshore waters before ocean currents and their ability to swim, transport them back to the coast. The abundance of western rock lobster puerulus (settlement phase post phyllosoma) has historically been observed to be positively correlated with the strength of the Leeuwin Current, and an index of puerulus numbers is used by fisheries managers as a predictor of subsequent lobster abundance 3–4 years later. In 2008 and 2009 the Leeuwin Current was strong, yet a settlement failure occurred throughout the fishery prompting management changes and a rethinking of environmental factors associated with their settlement. Thus, understanding factors that may have been responsible for the settlement failure is important for fisheries management. Oceanographic parameters likely to influence puerulus settlement were derived for a 17 year period to investigate correlations. Analysis indicated that puerulus settlement at adjacent monitoring sites have similar oceanographic forcing with kinetic energy in the offshore and the strength of the Leeuwin Current being key factors. Settlement failure years were synonymous with “hiatus” conditions in the south-east Indian Ocean, and periods of sustained cooler water present offshore. Post 2009, there has been an unusual but consistent increase in the Leeuwin Current during the early summer months with a matching decrease in the Capes Current, that may explain an observed settlement timing mismatch compared to historical data. Our study has revealed that a culmination of these conditions likely led to the recruitment failure and subsequent changes in puerulus settlement patterns.

Evaluation of otolith microstructures for ageing of larval white sturgeon

Sturgeon populations are endangered worldwide, mainly due to habitat degradation and overexploitation causing recruitment failure. Understanding of early development, survival and growth in sturgeon is limited by a lack of a validated method to directly estimate larval age. In a laboratory calibration study, we reared white sturgeon larvae from hatch for 3 weeks at 12 and 16°C and two feeding regimes (fed and unfed) to determine the usefulness of their vateritic micro-otoconia for larval ageing and the influence of environmental factors on ring structure and size. By marking the otoliths twice at known ages with Alizarin Red S, we were able to confirmed the presence of daily rings in the largest micro-otoconia and the feasibility of ageing larval sturgeon using otoliths. Three observers blind to age and treatment assessed larval age from daily rings with an overall precision of 67-82% and 30-70% accuracy, dependent on larval age. Neither temperature nor feeding had a significant effect on ring width or readability. Thus, ageing with otoliths in sturgeon is a promising tool in sturgeon conservation efforts.