An Immune-Related Gene Evolved into the Master Sex-Determining Gene in Rainbow Trout, Oncorhynchus mykiss

Since the discovery of Sry in mammals [1, 2], few other master sex-determining genes have been identified in vertebrates [3-7]. To date, all of these genes have been characterized as well-known factors in the sex differentiation pathway, suggesting that the same subset of genes have been repeatedly and independently selected throughout evolution as master sex determinants [8, 9]. Here, we characterized in rainbow trout an unknown gene expressed only in the testis, with a predominant expression during testicular differentiation. This gene is a male-specific genomic sequence that is colocalized along with the sex-determining locus. This gene, named sdY for sexually dimorphic on the Y?chromosome, encodes a protein that displays similarity to the C-terminal domain of interferon regulatory factor 9. The targeted inactivation of sdY in males using zinc-finger nuclease induces ovarian differentiation, and the overexpression of sdY in females using additive transgenesis induces testicular differentiation. Together, these results demonstrate that sdY is a novel vertebrate master sex-determining gene not related to any known sex-differentiating gene. These findings highlight an unexpected evolutionary plasticity in vertebrate sex determination through the demonstration that master sex determinants can arise from the de novo evolution of genes that have not been previously implicated in sex differentiation.     

How Much Do Metamemory Beliefs Contribute to the Font-Size Effect in Judgments of Learning?

Evidence shows that the font size of study items significantly influences judgments of learning (JOLs) and that people’s JOLs are generally higher for larger words than for smaller words. Previous studies have suggested that font size influences JOLs in a belief-based way. However, few studies have directly examined how much people’s beliefs contribute to the font-size effect in JOLs. This study investigated the degree to which font size influenced JOLs in a belief-based way. In Experiment 1, one group of participants (learners) studied words with different font sizes and made JOLs, whereas another group of participants (observers) viewed the learners'' study phase and made JOLs for the learners. In Experiment 2, participants made both JOLs and belief-based recall predictions for large and small words. Our results suggest that metamemory beliefs play an important role in the font-size effect in JOLs.     

Combined and Singular Effects of Dietary PrimaLac® and Potassium Diformate (KDF) on Growth Performance and Some Physiological Parameters of Rainbow Trout (Oncorhynchus mykiss)

The present study was conducted to assess the effects of combined and singular dietary administration of PrimaLac® and potassium diformate (KDF) on growth performance, feed utilization, digestive enzymes activity, and some physiological parameters of rainbow trout (Oncorhynchus mykiss) juvenile. Three hundred sixty rainbow trout juveniles (25 ± 1.8 g) were randomly stocked in 300-L tanks (30 fish/tank), and fed three times daily on a basal diet (control), diets incorporated with 12 g kg⁻¹ KDF (FT₁), 1.5 g kg⁻¹ PrimaLac® (FT₂), and combination of 1.5 g kg⁻¹ probiotic and 12 g kg⁻¹ KDF (FT₃) in triplicates, for 8 weeks. At the end of feeding trial, growth performance, body composition, digestive enzymes, liver enzymes, and biochemical parameters were measured. Our results revealed that combined administration of PrimaLac® and KDF (FT₃) exhibited significantly higher weight gain and specific growth rate (SGR) compared to other groups (P < 0.05). Glucose and cortisol levels showed no significant differences between fish fed different test diets (P > 0.05). The highest lipase, protease and amylase activity were observed in group of fish fed FT₃ followed by FT₂ and FT₁. Besides, the diets FT₂ and FT₃ led to significantly lower of ALP, ALT, and AST compared to control group. The present results indicated that combined administration of PrimaLac® and KDF can be considered as a beneficial feed additive and growth promotor for O. mykiss juvenile.

     

Protein Retention Assessment of Four Levels of Poultry By-Product Substitution of Fishmeal in Rainbow Trout (Oncorhynchus mykiss) Diets Using Stable Isotopes of Nitrogen (δ15N) as Natural Tracers

This is second part from an experiment where the nitrogen retention of poultry by-product meal (PBM) compared to fishmeal (FM) was evaluated using traditional indices. Here a quantitative method using stable isotope ratios of nitrogen (δ¹⁵N values) as natural tracers of nitrogen incorporation into fish biomass is assessed. Juvenile rainbow trout (Oncorhynchus mykiss) were fed for 80 days on isotopically distinct diets in which 0, 33, 66 and 100% of FM as main protein source was replaced by PBM. The diets were isonitrogenous, isolipidic and similar in gross energy content. Fish in all treatments reached isotopic equilibrium by the end of the experiment. Two-source isotope mixing models that incorporated the isotopic composition of FM and PBM as well as that of formulated feeds, empirically derived trophic discrimination factors and the isotopic composition of fish that had reached isotopic equilibrium to the diets were used to obtain a quantitative estimate of the retention of each source of nitrogen. Fish fed the diets with 33 and 66% replacement of FM by PBM retained poultry by-product meal roughly in proportion to its level of inclusion in the diets, whereas no differences were detected in the protein efficiency ratio. Coupled with the similar biomass gain of fishes fed the different diets, our results support the inclusion of PBM as replacement for fishmeal in aquaculture feeds. A re-feeding experiment in which all fish were fed a diet of 100% FM for 28 days indicated isotopic turnover occurred very fast, providing further support for the potential of isotopic ratios as tracers of the retention of specific protein sources into fish tissues. Stable isotope analysis is a useful tool for studies that seek to obtain quantitative estimates of the retention of different protein sources.     

Do sperm contribute to the buffering capacity of steelhead trout (Oncorhynchus mykiss) semen?

The motility of salmonid sperm is pH-sensitive and the buffering capacity of the seminal plasma is low. The objective of the present study was to determine the extent to which sperm contribute to the buffering capacity of whole steelhead (Oncorhynchus mykiss) semen. To determine the buffering capacity, semen and seminal plasma samples were titrated with HCl and pH measurements taken at 1-2 min. The buffering capacity of semen was not different from that of seminal plasma over the pH range 7.5 to 8.5 and was approximately 15% to 20% less over the range 6.0 to 7.0. Comparable results were obtained for the semen and seminal plasma of the chinook salmon (Oncorhynchus tshawytscha). To assess whether the intracellular environment could influence the buffering capacity, the effects of cell disruption with n-butanol and Triton X-100 (TX-100) were determined. Over the pH range 7.5 to 8.5, the presence of n-butanol or TX-100 resulted in a doubling of the buffering capacity of the semen; TX-100, but not n-butanol, increased semen buffering capacity over the pH range 6.0 to 7.0. To determine whether the sperm's intracellular compartment might contribute to the buffering capacity over a longer duration, semen and seminal plasma samples were acidified with HCl and the pH measured over several hours. These data suggest that intact sperm contribute no more than about 25% to the buffering capacity of whole semen. The buffering capacity of steelhead semen and seminal plasma were comparably and modestly temperature sensitive. The results suggest that the sperm may contribute to the buffering capacity of the semen over a physiological pH range, however, if so, the effect is relatively small.     

Deep Imaging: How Much of the Proteome Does Current Top-Down Technology Already Resolve?

Effective proteome analyses are based on interplay between resolution and detection. It had been claimed that resolution was the main factor limiting the use of two-dimensional gel electrophoresis. Improved protein detection now indicates that this is unlikely to be the case. Using a highly refined protocol, the rat brain proteome was extracted, resolved, and detected. In order to overcome the stain saturation threshold, high abundance protein species were excised from the gel following standard imaging. Gels were then imaged again using longer exposure times, enabling detection of lower abundance, less intensely stained protein species. This resulted in a significant enhancement in the detection of resolved proteins, and a slightly modified digestion protocol enabled effective identification by standard mass spectrometric methods. The data indicate that the resolution required for comprehensive proteome analyses is already available, can assess multiple samples in parallel, and preserve critical information concerning post-translational modifications. Further optimization of staining and detection methods promises additional improvements to this economical, widely accessible and effective top-down approach to proteome analysis.     

Does Biofilm Contribute to Diel Cycling of Zn in High Ore Creek, Montana?

Concentrations of metals cycle daily in the water column of some mining-impacted streams in the Rocky Mountains of the western USA. We hypothesized that biofilm in High Ore Creek, Montana, USA, sorbs and releases Zn on a diel cycle, and this uptake-and-release cycle controls the total and dissolved (0.45-μm filtered) Zn concentrations. We collected water samples from three sites (upstream, middle and downstream at 0, 350 and 650 m, respectively) along a 650-m reach of High Ore Creek during a 47-h period in August 2002 and from the upstream and downstream sites during a 24-h period in August 2003; we also collected biofilm samples at these sites. In 2002 and 2003, total and dissolved Zn concentrations did not exhibit a diel cycle at the upstream sampling site, which was ~30 m downstream from a settling pond through which the creek flows. However, total and dissolved Zn concentrations exhibited a diel cycle at the middle and downstream sampling sites, with the highest Zn concentrations occurring at dawn and the lowest Zn concentrations occurring during late afternoon (>2-fold range of concentrations at the downstream site). Based on (1) concentrations of Zn in biofilm at the three sites and (2) results of streamside experiments that demonstrated Zn uptake and release by na?ve biofilm during the light and dark hours of a photocycle, respectively, we conclude that Zn uptake in photosynthetic biofilms could contribute a large percentage to the cycling of Zn concentrations in the water column in High Ore Creek.     

How Much Does It Cost to Improve Access to Voluntary Medical Male Circumcision among High-Risk,Low-Income Communities in Uganda?

Background

The Ugandan Ministry of Health has endorsed voluntary medical male circumcision as an HIV prevention strategy and has set ambitious goals (e.g., 4.2 million circumcisions by 2015). Innovative strategies to improve access for hard to reach, high risk, and poor populations are essential for reaching such goals. In 2009, the Makerere University Walter Reed Project began the first facility-based VMMC program in Uganda in a non-research setting. In addition, a mobile clinic began providing VMMC services to more remote, rural locations in 2011. The primary objective of this study was to estimate the average cost of performing VMMCs in the mobile clinic compared to those performed in health facilities (fixed sites). The difference between such costs is the cost of improving access to VMMC.

Methods

A micro-costing approach was used to estimate costs from the service provider’s perspective of a circumcision. Supply chain and higher-level program support costs are not included.

Results

The average cost (US$2012) of resources used per circumcision was $61 in the mobile program ($72 for more remote locations) compared to $34 at the fixed site. Costs for community mobilization, HIV testing, the initial medical exam, and staff for performing VMMC operations were similar for both programs. The cost of disposable surgical kits, the additional upfront cost for the mobile clinic, and additional costs for staff drive the differences in costs between the two programs. Cost estimates are relatively insensitive to patient flow over time.

Conclusion

The MUWRP VMMC program improves access for hard to reach, relatively poor, and high-risk rural populations for a cost of $27-$38 per VMMC. Costs to patients to access services are almost certainly less in the mobile program, by reducing out-of-pocket travel expenses and lost time and associated income, all of which have been shown to be barriers for accessing treatment.     

Reaction Norms in Natural Conditions: How Does Metabolic Performance Respond to Weather Variations in a Small Endotherm Facing Cold Environments?

Reaction norms reflect an organisms'' capacity to adjust its phenotype to the environment and allows for identifying trait values associated with physiological limits. However, reaction norms of physiological parameters are mostly unknown for endotherms living in natural conditions. Black-capped chickadees (Poecile atricapillus) increase their metabolic performance during winter acclimatization and are thus good model to measure reaction norms in the wild. We repeatedly measured basal (BMR) and summit (Msum) metabolism in chickadees to characterize, for the first time in a free-living endotherm, reaction norms of these parameters across the natural range of weather variation. BMR varied between individuals and was weakly and negatively related to minimal temperature. Msum varied with minimal temperature following a Z-shape curve, increasing linearly between 24°C and −10°C, and changed with absolute humidity following a U-shape relationship. These results suggest that thermal exchanges with the environment have minimal effects on maintenance costs, which may be individual-dependent, while thermogenic capacity is responding to body heat loss. Our results suggest also that BMR and Msum respond to different and likely independent constraints.     

Understanding the Causes of Recent Warming of Mediterranean Waters. How Much Could Be Attributed to Climate Change?

During the past two decades, Mediterranean waters have been warming at a rather high rate resulting in scientific and social concern. This warming trend is observed in satellite data, field data and model simulations, and affects both surface and deep waters throughout the Mediterranean basin. However, the warming rate is regionally different and seems to change with time, which has led to the question of what causes underlie the observed trends. Here, we analyze available satellite information on sea surface temperature (SST) from the last 25 years using spectral techniques and find that more than half of the warming tendency during this period is due to a non-linear, wave-like tendency. Using a state of the art hydrodynamic model, we perform a hindcast simulation and obtain the simulated SST evolution of the Mediterranean basin for the last 52 years. These SST results show a clear sinusoidal tendency that follows the Atlantic Multidecadal Oscillation (AMO) during the simulation period. Our results reveal that 58% of recent warming in Mediterranean waters could be attributed to this AMO-like oscillation, being anthropogenic-induced climate change only responsible for 42% of total trend. The observed acceleration of water warming during the 1990s therefore appears to be caused by a superimposition of anthropogenic-induced warming with the positive phase of the AMO, while the recent slowdown of this tendency is likely due to a shift in the AMO phase. It has been proposed that this change in the AMO phase will mask the effect of global warming in the forthcoming decades, and our results indicate that the same could also be applicable to the Mediterranean Sea. Henceforth, natural multidecadal temperature oscillations should be taken into account to avoid underestimation of the anthropogenic-induced warming of the Mediterranean basin in the future.     

Does Virus-Induced Lysis Contribute Significantly to Bacterial Mortality in the Oxygenated Sediment Layer of Shallow Oxbow Lakes?

Despite the recognition that viruses are ubiquitous components of aquatic ecosystems, the number of studies on viral abundance and the ecological role of viruses in sediments is scarce. In this investigation, the interactions between viruses and bacteria were studied in the oxygenated silty sediment layer of a mesotrophic oxbow lake. A long-term study (13 months) and a diel study revealed that viruses are a numerically important and dynamic component of the microbial community. The abundance and decay rates ranged from 4.3 × 10⁹ to 7.2 × 10⁹ particles ml of wet sediment⁻¹ and from undetectable to 22.2 × 10⁷ particles ml⁻¹ h⁻¹, respectively, and on average the values were 2 orders of magnitude higher than the values for the overlying water. In contrast to our expectations, viruses did not contribute significantly to the bacterial mortality in the sediment, since on average only 6% (range, 0 to 25%) of the bacterial secondary production was controlled by viruses. The low impact of viruses on the bacterial community may be associated with the quantitatively low viral burden that benthic bacteria have to cope with compared to the viral burden with which bacterial assemblages in the water column are confronted. The virus-to-bacterium ratio of the sediment varied between 0.9 and 3.2, compared to a range of 5.0 to 12.4 obtained for the water column. We speculate that despite high numbers of potential hosts, the possibility of encountering a host cell is limited by the physical conditions in the sediment, which is therefore not a favorable environment for viral proliferation. Our data suggest that viruses do not play an important role in the processing and transfer of bacterial carbon in the oxygenated sediment layer of the environment investigated.     

How Does Conversion of Natural Tropical Rainforest Ecosystems Affect Soil Bacterial and Fungal Communities in the Nile River Watershed of Uganda?

Uganda''s forests are globally important for their conservation values but are under pressure from increasing human population and consumption. In this study, we examine how conversion of natural forest affects soil bacterial and fungal communities. Comparisons in paired natural forest and human-converted sites among four locations indicated that natural forest soils consistently had higher pH, organic carbon, nitrogen, and calcium, although variation among sites was large. Despite these differences, no effect on the diversity of dominant taxa for either bacterial or fungal communities was detected, using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Composition of fungal communities did generally appear different in converted sites, but surprisingly, we did not observe a consistent pattern among sites. The spatial distribution of some taxa and community composition was associated with soil pH, organic carbon, phosphorus and sodium, suggesting that changes in soil communities were nuanced and require more robust metagenomic methods to understand the various components of the community. Given the close geographic proximity of the paired sampling sites, the similarity between natural and converted sites might be due to continued dispersal between treatments. Fungal communities showed greater environmental differentiation than bacterial communities, particularly according to soil pH. We detected biotic homogenization in converted ecosystems and substantial contribution of β-diversity to total diversity, indicating considerable geographic structure in soil biota in these forest communities. Overall, our results suggest that soil microbial communities are relatively resilient to forest conversion and despite a substantial and consistent change in the soil environment, the effects of conversion differed widely among sites. The substantial difference in soil chemistry, with generally lower nutrient quantity in converted sites, does bring into question, how long this resilience will last.     

How Much Does it Cost to Expand a Protected Area System? Some Critical Determining Factors and Ranges of Costs for Queensland

Many governments have recently gone on record promising large-scale expansions of protected areas to meet global commitments such as the Convention on Biological Diversity. As systems of protected areas are expanded to be more comprehensive, they are more likely to be implemented if planners have realistic budget estimates so that appropriate funding can be requested. Estimating financial budgets a priori must acknowledge the inherent uncertainties and assumptions associated with key parameters, so planners should recognize these uncertainties by estimating ranges of potential costs. We explore the challenge of budgeting a priori for protected area expansion in the face of uncertainty, specifically considering the future expansion of protected areas in Queensland, Australia. The government has committed to adding ～12 million ha to the reserve system, bringing the total area protected to 20 million ha by 2020. We used Marxan to estimate the costs of potential reserve designs with data on actual land value, market value, transaction costs, and land tenure. With scenarios, we explored three sources of budget variability: size of biodiversity objectives; subdivision of properties; and legal acquisition routes varying with tenure. Depending on the assumptions made, our budget estimates ranged from $214 million to $2.9 billion. Estimates were most sensitive to assumptions made about legal acquisition routes for leasehold land. Unexpected costs (costs encountered by planners when real-world costs deviate from assumed costs) responded non-linearly to inability to subdivide and percentage purchase of private land. A financially conservative approach--one that safeguards against large cost increases while allowing for potential financial windfalls--would involve less optimistic assumptions about acquisition and subdivision to allow Marxan to avoid expensive properties where possible while meeting conservation objectives. We demonstrate how a rigorous analysis can inform discussions about the expansion of systems of protected areas, including the identification of factors that influence budget variability.     

Does Ownership Matter? An Overview of Systematic Reviews of the Performance of Private For-Profit,Private Not-For-Profit and Public Healthcare Providers

Introduction

Ownership of healthcare providers has been considered as one factor that might influence their health and healthcare related performance. The aim of this article was to provide an overview of what is known about the effects on economic, administrative and health related outcomes of different types of ownership of healthcare providers -namely public, private non-for-profit (PNFP) and private for-profit (PFP)- based on the findings of systematic reviews (SR).

Methods and Findings

An overview of systematic reviews was performed. Different databases were searched in order to select SRs according to an explicit comprehensive criterion. Included SRs were assessed to determine their methodological quality. Of the 5918 references reviewed, fifteen SR were included, but six of them were rated as having major limitations, so they weren''t incorporated in the analyses. According to the nine analyzed SR, ownership does seem to have an effect on health and healthcare related outcomes. In the comparison of PFP and PNFP providers, significant differences in terms of mortality of patients and payments to facilities have been found, both being higher in PFP facilities. In terms of quality and economic indicators such as efficiency, there are no concluding results. When comparing PNFP and public providers, as well as for PFP and public providers, no clear differences were found.

Conclusion

PFP providers seem to have worst results than their PNFP counterparts, but there are still important evidence gaps in the literature that needs to be covered, including the comparison between public and both PFP and PNFP providers. More research is needed in low and middle income countries to understand the impact on and development of healthcare delivery systems.     

Interventional spine procedures under CT guidance: How to reduce patient radiation dose without compromising the successful outcome of the procedure?

This technical note proposes a method to reduce radiation dose for spine interventions under CT guidance without compromising the successful outcome of the procedure. Two consecutive periods of 14 months before and after optimization were investigated with 162 and 440 patients, respectively. By optimizing the acquisition parameters (decreased kV and mAs) and appropriately adjusting the reconstruction (kernels, slice thickness, etc) and visualization parameters, image quality was maintained suitable to perform the procedure. By reducing both kV and mAs, dose was reduced by 72% on fluoroscopy mode (i-fluoro) and sequential mode (i-sequence). Moreover, dose was reduced by 58% on helical mode (i-spiral). Depending on the radiologist, the fluoroscopy time was decreased by between 37% and 56%. Acquisitions with i-sequence were less irradiating than the i-fluoro or the i-spiral modes. Radiation doses were reduced by 65% for infiltrations, 51% for vertebral expansions, and 56% for bone biopsies. Median (1st quartile; 3rd quartile) effective dose were 2.1 (1.3; 3.5) mSv, 10.8 (6.7; 18.3) mSv for and 3.0 (2.4; 4.3) mSv, respectively. Radiologists reported “satisfactory” image quality. During interventional spine procedures under CT scan, reducing kV and mAs associated with the use of i-sequence substantially reduces patient dose.     

How to Handle Speciose Clades? Mass Taxon-Sampling as a Strategy towards Illuminating the Natural History of Campanula (Campanuloideae)

Background

Speciose clades usually harbor species with a broad spectrum of adaptive strategies and complex distribution patterns, and thus constitute ideal systems to disentangle biotic and abiotic causes underlying species diversification. The delimitation of such study systems to test evolutionary hypotheses is difficult because they often rely on artificial genus concepts as starting points. One of the most prominent examples is the bellflower genus Campanula with some 420 species, but up to 600 species when including all lineages to which Campanula is paraphyletic. We generated a large alignment of petD group II intron sequences to include more than 70% of described species as a reference. By comparison with partial data sets we could then assess the impact of selective taxon sampling strategies on phylogenetic reconstruction and subsequent evolutionary conclusions.

Methodology/Principal Findings

Phylogenetic analyses based on maximum parsimony (PAUP, PRAP), Bayesian inference (MrBayes), and maximum likelihood (RAxML) were first carried out on the large reference data set (D680). Parameters including tree topology, branch support, and age estimates, were then compared to those obtained from smaller data sets resulting from “classification-guided” (D088) and “phylogeny-guided sampling” (D101). Analyses of D088 failed to fully recover the phylogenetic diversity in Campanula, whereas D101 inferred significantly different branch support and age estimates.

Conclusions/Significance

A short genomic region with high phylogenetic utility allowed us to easily generate a comprehensive phylogenetic framework for the speciose Campanula clade. Our approach recovered 17 well-supported and circumscribed sub-lineages. Knowing these will be instrumental for developing more specific evolutionary hypotheses and guide future research, we highlight the predictive value of a mass taxon-sampling strategy as a first essential step towards illuminating the detailed evolutionary history of diverse clades.