Individual Pools Research Articles

Evidence for transovarial transmission of tick-borne rickettsiae circulating in Northern Mongolia.

Transstadial transmission of tick-borne rickettsiae has been well documented. Few studies, however, have evaluated the role of transovarial transmission of tick-borne rickettsiae, particularly in nature within the host-vector ecosystem. This cross-sectional study aimed to understand the role of transovarial transmission of tick-borne rickettsiae among feeding ticks at different life stages. Tick eggs laid by engorged wild-caught adult female ticks were pooled and tested for Rickettsia spp. and Anaplasma/Ehrlichia spp. using molecular techniques, while adult fed ticks were tested individually. Additionally, larval and nymphal ticks were collected in the wild from small mammals, pooled and tested for Rickettsia spp. and Anaplasma/Ehrlichia spp. There were 38 fed adult and 618 larvae/nymphs (60 pools total) Dermacentor spp. ticks collected from livestock and rodents. All individual adult ticks and tick pools were positive for Rickettsia spp. While none of the larvae/nymphs were positive for Anaplasma/Ehrlichia spp., two adult fed ticks were positive. Rickettsia spp. DNA was detected in 91% (30/33) of the pooled eggs tested, and one pool of eggs tested positive for Anaplasma/Ehrlichia spp. Sequencing data revealed Rickettsia spp. shared ≥99% identity with R. raoultii ompA. Anaplasma/Ehrlichia spp. shared ≥89% identity with A. ovis 16S ribosomal RNA. This study identified potential transovarial transmission of Rickettsia spp. and Anaplasma spp. among D. nuttalli ticks. Additional studies are needed to further assess the proportion of transovarial transmission occurring in nature to better understand the burden and disease ecology of tick-borne rickettsiae in Mongolia.

A radiative forcing analysis of tropical peatlands before and after their conversion to agricultural plantations.

The tropical peat swamp forests of South-East Asia are being rapidly converted to agricultural plantations of oil palm and Acacia creating a significant global "hot-spot" for CO2 emissions. However, the effect of this major perturbation has yet to be quantified in terms of global warming potential (GWP) and the Earth's radiative budget. We used a GWP analysis and an impulse-response model of radiative forcing to quantify the climate forcing of this shift from a long-term carbon sink to a net source of greenhouse gases (CO2 and CH4 ). In the GWP analysis, five tropical peatlands were sinks in terms of their CO2 equivalent fluxes while they remained undisturbed. However, their drainage and conversion to oil palm and Acacia plantations produced a dramatic shift to very strong net CO2 -equivalent sources. The induced losses of peat carbon are ~20× greater than the natural CO2 sequestration rates. In contrast, a radiative forcing model indicates that the magnitude of this shift from a net cooling to warming effect is ultimately related to the size of an individual peatland's carbon pool. The continuous accumulation of carbon in pristine tropical peatlands produced a progressively negative radiative forcing (i.e., cooling) that ranged from -2.1 to -6.7nW/m2 per hectare peatland by 2010 CE, referenced to zero at the time of peat initiation. Peatland conversion to plantations leads to an immediate shift from negative to positive trend in radiative forcing (i.e., warming). If drainage persists, peak warming ranges from +3.3 to +8.7nW/m2 per hectare of drained peatland. More importantly, this net warming impact on the Earth's radiation budget will persist for centuries to millennia after all the peat has been oxidized to CO2 . This previously unreported and undesirable impact on the Earth's radiative balance provides a scientific rationale for conserving tropical peatlands in their pristine state.

Limited nitrate retention capacity in the Upper Mississippi River

The Mississippi River and other large rivers have the potential to regulate nitrogen export from terrestrial landscapes, and thus mitigate eutrophication in downstream aquatic ecosystems. In large rivers, human-constructed impoundments and connected backwaters may facilitate nitrogen removal; however, the capacity of these features is poorly quantified and incompletely incorporated into model frameworks. Using a high-resolution and spatially intensive sampling technique, we assessed the contribution of individual navigation pools, as well as impounded open waters and backwater wetlands within them, to overall nitrate retention by mapping the entire length (1370 km) of the Upper Mississippi River (UMR) main channel. Based on this single spatial survey of water chemistry, the river appeared to act primarily as a passive nitrate transporter, retaining only 12.5% of the incoming load, most of which occurred in the upper 150 km of the river, which includes the largest and only naturally impounded reach of the river. Although reservoirs typically are nitrogen sinks, our data indicate that UMR dams do not impede river flows to the extent necessary to promote substantial changes in water residence times and subsequent nitrogen removal. Backwaters routinely had lower nitrate concentrations than the main channel, but their limited hydrologic connectivity to the through-flowing river channel constrained their influence on downstream export. As a whole, the UMR did not remove a substantial proportion of its nitrate load despite optimal N removal conditions, numerous impoundments, and the presence of extensive backwater habitats. These results suggest that efforts to reduce delivery of nitrogen to the Gulf of Mexico should emphasize mitigation strategies that target upland nutrient sources rather than relying on removal within the Mississippi River.

Implications of plasma thiol redox in disease.

Thiol groups are crucially involved in signaling/homeostasis through oxidation, reduction, and disulphide exchange. The overall thiol pool is the resultant of several individual pools of small compounds (e.g. cysteine), peptides (e.g. glutathione), and thiol proteins (e.g. thioredoxin (Trx)), which are not in equilibrium and present specific oxidized/reduced ratios. This review addresses mechanisms and implications of circulating plasma thiol/disulphide redox pools, which are involved in several physiologic processes and explored as disease biomarkers. Thiol pools are regulated by mechanisms linked to their intrinsic reactivity against oxidants, concentration of antioxidants, thiol-disulphide exchange rates, and their dynamic release/removal from plasma. Major thiol couples determining plasma redox potential (Eh) are reduced cysteine (CyS)/cystine (the disulphide form of cysteine) (CySS), followed by GSH/disulphide-oxidized glutathione (GSSG). Hydrogen peroxide and hypohalous acids are the main plasma oxidants, while water-soluble and lipid-soluble small molecules are the main antioxidants. The thiol proteome and thiol-oxidoreductases are emerging investigative areas given their specific disease-related responses (e.g. protein disulphide isomerases (PDIs) in thrombosis). Plasma cysteine and glutathione redox couples exhibit pro-oxidant changes directly correlated with ageing/age-related diseases. We further discuss changes in thiol-disulphide redox state in specific groups of diseases: cardiovascular, cancer, and neurodegenerative. These results indicate association with the disease states, although not yet clear-cut to yield specific biomarkers. We also highlight mechanisms whereby thiol pools affect atherosclerosis pathophysiology. Overall, it is unlikely that a single measurement provides global assessment of plasma oxidative stress. Rather, assessment of individual thiol pools and thiol-proteins specific to any given condition has more solid and logical perspective to yield novel relevant information on disease risk and prognosis.

Acquisition of Bartonella elizabethae by Experimentally Exposed Oriental Rat Fleas (Xenopsylla cheopis; Siphonaptera, Pulicidae) and Excretion of Bartonella DNA in Flea Feces.

Few studies have been able to provide experimental evidence of the ability of fleas to maintain rodent-associated Bartonella infections and excrete these bacteria. These data are important for understanding the transmission cycles and prevalence of these bacteria in hosts and vectors. We used an artificial feeding approach to expose groups of the oriental rat flea (Xenopsylla cheopis Rothschild; Siphonaptera, Pulicidae) to rat blood inoculated with varying concentrations of Bartonella elizabethae Daly (Bartonellaceae: Rhizobiales). Flea populations were maintained by membrane feeding on pathogen-free bloodmeals for up to 13 d post infection. Individual fleas and pools of flea feces were tested for the presence of Bartonella DNA using molecular methods (quantitative and conventional polymerase chain reaction [PCR]). The threshold number of Bartonellae required in the infectious bloodmeal for fleas to be detected as positive was 106 colony-forming units per milliliter (CFU/ml). Individual fleas were capable of harboring infections for at least 13 d post infection and continuously excreted Bartonella DNA in their feces over the same period. This experiment demonstrated that X. cheopis are capable of acquiring and excreting B. elizabethae over several days. These results will guide future work to model and understand the role of X. cheopis in the natural transmission cycle of rodent-borne Bartonella species. Future experiments using this artificial feeding approach will be useful for examining the horizontal transmission of B. elizabethae or other rodent-associated Bartonella species to naïve hosts and for determining the viability of excreted bacteria.

Combined influence of meteoric water and protein intake on hydrogen isotope values in archaeological human bone collagen

Hydrogen isotopes in archaeological human bone collagen are poorly understood, but present an opportunity to add new depth to our understanding of ancient populations. The competing influences of meteoric water versus protein intake on human bone collagen hydrogen isotope values were examined through comparison with the well-understood proxies of hydroxyapatite oxygen and collagen nitrogen isotopes, respectively. Consideration of the data set as individual points compared to averaged pools of individuals in each of 11 archaeological sites suggested the latter partially eliminates inherent variability due to food choice or regional movement. Collagen hydrogen isotopes were moderately correlated with hydroxyapatite oxygen isotopes (R = 0.695, site averages) and collagen nitrogen isotopes (R = 0.562, site averages). Correlation improved with a multiple linear regression including both oxygen and nitrogen (R = 0.745, site averages). Correlation between meteoric water hydrogen and oxygen isotope values converted from hydroxyapatite and collagen values, respectively, yielded a slope well below the expected value of ∼8 observed directly in meteoric water (i.e. the “meteoric water line”). Correlation between converted meteoric water hydrogen and the measured collagen non-exchangeable hydrogen isotope values showed a slope well below the expected value of 1.0. Theoretical meteoric water hydrogen isotope values and theoretical herbivorous collagen hydrogen isotope values were calculated based on previously established equations in order to construct a hypothetical framework free of trophic level influences. Deviations between actual values and these theoretical values correlated weakly with collagen nitrogen isotope values, suggesting that direct trophic level enrichment/depletion is not controlling the disparity between expected and measured values. The deviations are hypothetically caused by non-local food sources, and a decoupling of expected oxygen and hydrogen relationships as individuals consumed more meat and decreased in vivo non-essential amino acid production. This work presents a new model that facilitates understanding of the complex relationship between meteoric water and protein intake controls on hydrogen isotopes in omnivorous human populations that can potentially inform about past meteoric water values and amounts of animal protein consumption.

A genetic analysis of the European hedgehog (Erinaceus europaeus): an applicative case study to support its eradication from Pianosa Island (Tuscan Archipelago)

The introduction of allochthonous species represents a serious threat for the native gene pools and ecosystem biodiversity. The effect is particularly disastrous for insular biocoenoses, such as in the Tuscan archipelago, one of the most important biodiversity hotspot in the Mediterranean area. The EU tool LIFE + has funded an eradication project involving a set of allochthonous species on Pianosa Island ( http://www.restoconlife.eu ), including the European hedgehog (Erinaceus europaeus). Since eradication projects should not leave out of consideration a genetic analysis of the target species, the aim of our study was to characterize the genetic profile of the Pianosa hedgehog population. In particular, the data obtained had to help assessing the most compatible area for the release of all captured individuals. In the present work, eleven microsatellite loci and two mitochondrial gene portions (COXI and 16S) were characterized in individuals of E. europaeus from Pianosa, Elba, Sardinia Islands and mainland Italy. Both mtDNA and microsatellite data confirmed that the present-day population of Pianosa has an extremely low genetic diversity and a profile very similar to that of Elba. Consequently, our results do suggest that the Pianosa hedgehogs originated from a pool of individuals moved by human from Elba in recent times and could be relocated there.

Temperature affects phenological synchrony in a tree-killing bark beetle.

Phenological synchrony can promote population growth in species with positive density dependence. Variation among life stages in the thermal thresholds for development can foster phenological synchrony under thermal regimes that include frequent occurrence of temperatures between developmental thresholds. The southern pine beetle is an insect with positive density dependence that has recently undergone important shifts in population abundance at the northern extremes of their distribution. We evaluated the hypothesis that cooler winter temperatures in their northern range cause a convergence of the population life stage structure that leads to synchrony in spring flight phenology. We used a combination of approaches. First, in situ laboratory experiments demonstrated a threshold temperature for pupation that was greater than was required for larval development; rearing larvae at lower temperatures increased the pooling of individuals at the end stage of larval development and synchrony in adult emergence. Second, a development rate model showed a similar convergence of the majority of the population at the end stage of larval development when brood experienced the cooler temperatures of the northern region, but not with temperatures from the southern region, or as a null model. Finally, field trapping of wild beetles showed greater synchrony in the pine forests of New Jersey than in the warmer, historically occupied forests of Georgia and Mississippi. Given these results, pine-dominated forests in the northern edge of the southern pine beetle's range may experience more frequent occurrence of outbreaks, due to the positive feedbacks associated with a synchronous spring emergence of this insect.

Modelling credit and savings behaviour of chit fund participants

Chit funds are a form of Rotating Saving and Credit Association (ROSCA) prevalent in India. In this institution, a group of individuals pool in equal amounts of money at a fixed frequency, and at every time period a round of competitive bidding takes place among the individuals to identify a borrower for the collected amount. The borrower foregoes his/her right to participate in further auctions, but continues paying his/her share of the pool till every individual in the group has collected the pooled amount. The auction process relies on the bidders’ willingness to give up a certain amount of the pool and take the rest as loan. The amount given up at every time period is shared equally among all individuals in the group. A chit fund, therefore, is structurally similar to the modern formal banking and financial institutions: they act as an intermediary to optimally mobilize funds collected from savers to borrowers and manage repayment of loans from borrowers such that savers receive their dues at the appropriate time. The latter role exposes the industry to credit risk. Modern banking and financial institutions rely heavily on their ability to assess and mitigate credit risk; and, over the last century, they have been able to move from a system where risk assessment was based on human judgment and simple intuition to a system reliant on statistical and mathematical techniques. However, the standard chit fund company still relies heavily on human judgment for assessing risk. The pilot envisages a move towards a standardized statistical methodology for risk assessment and mitigation as the path ahead for the industry.

Utility of pooled sequencing for association mapping in nonmodel organisms.

High-density genome-wide sequencing increases the likelihood of discovering genes of major effect and genomic structural variation in organisms. While there is an increasing availability of reference genomes across broad taxa, the greatest limitation to whole-genome sequencing of multiple individuals continues to be the costs associated with sequencing. To alleviate excessive costs, pooling multiple individuals with similar phenotypes and sequencing the homogenized DNA (Pool-Seq) can achieve high genome coverage, but at the loss of individual genotypes. Although Pool-Seq has been an effective method for association mapping in model organisms, it has not been frequently utilized in natural populations. To extend bioinformatic tools for rapid implementation of Pool-Seq data in nonmodel organisms, we developed a pipeline called PoolParty and illustrate its effectiveness in genetic association mapping. Alignment expectations based on five pooled Chinook salmon (Oncorhynchus tshawytscha) libraries showed that approximately 48% genome coverage per library could be achieved with reasonable sequencing effort. We additionally examined male and female O.tshawytscha libraries to illustrate how Pool-Seq techniques can successfully map known genes associated with functional differences among sexes such as growth hormone 2. Finally, we compared pools of individuals of different spawning ages for each sex to discover novel genes involved with age at maturity in O.tshawytscha such as opsin4 and transmembrane protein19. While not appropriate for every system, Pool-Seq data processed by the PoolParty pipeline is a practical method for identifying genes of major effect in nonmodel organisms when high genome coverage is necessary and cost is a limiting factor.

Genetic characterization of a novel picorna-like virus in Culex spp. mosquitoes from Mozambique

BackgroundMosquitoes are the potential vectors for a variety of viruses that can cause diseases in the human and animal populations. Viruses in the order Picornavirales infect a broad range of hosts, including mosquitoes. In this study, we aimed to characterize a novel picorna-like virus from the Culex spp. of mosquitoes from the Zambezi Valley of Mozambique.MethodsThe extracted RNA from mosquito pools was pre-amplified with the sequence independent single primer amplification (SISPA) method and subjected to high-throughput sequencing using the Ion Torrent platform. Reads that are classified as Iflaviridae, Picornaviridae and Dicistroviridae were assembled by CodonCode Aligner and SPAdes. Gaps between the viral contigs were sequenced by PCR. The genomic ends were analyzed by 5′ and 3′ RACE PCRs. The ORF was predicted with the NCBI ORF finder. The conserved domains were identified with ClustalW multiple sequence alignment, and a phylogenetic tree was built with MEGA. The presence of the virus in individual mosquito pools was detected by RT-PCR assay.ResultsA near full-length viral genome (9740 nt) was obtained in Culex mosquitoes that encoded a complete ORF (3112 aa), named Culex picorna-like virus (CuPV-1). The predicted ORF had 38% similarity to the Hubei picorna-like virus 35. The sequence of the conserved domains, Helicase-Protease-RNA-dependent RNA polymerase, were identified by multiple sequence alignment and found to be at the 3′ end, similar to iflaviruses. Phylogenetic analysis of the putative RdRP amino acid sequences indicated that the virus clustered with members of the Iflaviridae family. CuPV-1 was detected in both Culex and Mansonia individual pools with low infection rates.ConclusionsThe study reported a highly divergent, near full-length picorna-like virus genome from Culex spp. mosquitoes from Mozambique. The discovery and characterization of novel viruses in mosquitoes is an initial step, which will provide insights into mosquito-virus interaction mechanisms, genetic diversity and evolution.

The ecology of a system of natural mesocosms: Rock pools in the Atlantic Forest

Abstract The methodological trade‐off between the realism of natural systems and the tractability of artificial systems has led ecologists to praise the qualities of natural micro‐ and mesocosms as an ideal means for investigating the processes that organise biological communities. Among the different types of naturally occurring micro‐ and mesocosms, clusters of rock pools combine a global distribution with the simplicity, replicability, tractability and structural homogeneity that are desirable in model experimental systems. However, critical geographical data gaps in the study of rock pools must be filled if ecologists are to use these as global model systems in community ecology. In a year‐long study of a cluster of 181 rock pools in the Atlantic Forest of eastern Brazil, we conducted the first study on the ecology of freshwater rock pool communities in the entire Neotropical region, to the best of our knowledge. Building on published descriptions of the factors that drive community assembly within rock pools, we tested the hypotheses (1) that the effects of environmental factors prevail over those of spatial factors and (2) that pool volume and secondarily species interactions between consumers (anuran larvae), resources (leaf litter) and predators (dragonfly larvae) have a dominant role in driving species patterns of occurrence and abundance. Our hypotheses were generally supported as community structure was clearly influenced by the environmental characteristics of individual pools, and not to the spatial relations among them. Pool volume had a consistent, positive influence on the probability of occurrence of dragonflies and tadpoles of three anuran species, possibly through the influence of volume on hydroperiod. However, the species most strongly associated with larger pools were not those with longer times until metamorphosis, but those with larger egg clutches. In addition, species occurrences were positively associated with resource availability as measured by leaf litter mass (significantly so for Rhinella and dragonflies). These observations suggest that within these severely confined freshwater systems, indicators of environmental stability and reduced intraspecific competition could be important ultimate criteria for oviposition site selection. The ease of sampling and manipulating the rock pools at our Atlantic Forest site combined with the observation that these pools are governed by some of the same factors influencing community organisation in other rock pool systems around the world reinforces the merit of the system as a means to conduct global comparative studies of the ecology of communities and metacommunities.

Sialome diversity of ticks revealed by RNAseq of single tick salivary glands.

Ticks salivate while feeding on their hosts. Saliva helps blood feeding through host anti-hemostatic and immunomodulatory components. Previous transcriptomic and proteomic studies revealed the complexity of tick saliva, comprising hundreds of polypeptides grouped in several multi-genic families such as lipocalins, Kunitz-domain containing peptides, metalloproteases, basic tail secreted proteins, and several other families uniquely found in ticks. These studies also revealed that the composition of saliva changes with time; expression of transcripts from the same family wax and wane as a function of feeding time. Here, we examined whether host immune factors could influence sialome switching by comparing sialomes of ticks fed naturally on a rabbit, to ticks artificially fed on defibrinated blood depleted of immune components. Previous studies were based on transcriptomes derived from pools of several individuals. To get an insight into the uniqueness of tick sialomes, we performed transcriptomic analyses of single salivary glands dissected from individual adult female I. ricinus ticks. Multivariate analysis identified 1,279 contigs differentially expressed as a function of time and/or feeding mode. Cluster analysis of these contigs revealed nine clusters of differentially expressed genes, four of which appeared consistently across several replicates, but five clusters were idiosyncratic, pointing to the uniqueness of sialomes in individual ticks. The disclosure of tick quantum sialomes reveals the unique salivary composition produced by individual ticks as they switch their sialomes throughout the blood meal, a possible mechanism of immune evasion.

Carbon fluxes and interannual drivers in a temperate forest ecosystem assessed through comparison of top-down and bottom-up approaches

Despite decades of research, gaining a comprehensive understanding of carbon (C) cycling in forests remains a considerable challenge. Uncertainties stem from persistent methodological limitations and the difficulty of resolving top-down estimates of ecosystem C exchange with bottom-up measurements of individual pools and fluxes. To address this, we derived estimates and associated uncertainties of ecosystem C fluxes for a 100–125 year old mixed temperate forest stand at the Bartlett Experimental Forest, New Hampshire, USA, using three different approaches: (1) tower-based eddy covariance, (2) a biometric approach involving C flux measurements of individual ecosystem subcomponents, and (3) an inventory approach involving changes in major C stocks over time. Our analysis made use of 13 years of data, collected over the period from 2004 to 2016.Estimates of mean annual net ecosystem production (NEP) ranged from 120 to 133 g C m−2, demonstrating strong agreement among methods and suggesting that this aging forest acts as a moderate C sink. The use of multiple approaches to measure C fluxes and their uncertainties helped place constraints on difficult-to-measure processes such as aboveground contributions to ecosystem respiration and belowground allocation to mycorrhizal fungal biomass (which was estimated at 20% of net primary production).Analysis of interannual variability in C fluxes revealed a decoupling between annual wood growth and either current year or lagged NEP or GPP, suggesting that source limitation (C supply) is likely not controlling rates of wood production, at least on an interannual scale. Results also demonstrated a strong association between the maximum rate of C uptake during the growing season (Amax) and the length of the vernal window, defined as the period of time between soil thaw and the onset of photosynthesis. This suggests an important, but poorly understood, influence of winter and spring climate on mid-summer canopy physiology. Efforts to resolve the mechanisms responsible should be prioritized in light of ongoing and predicted changes in climate for the northeastern U.S. region, particularly during the winter and winter-spring transition period.

Functional circuit architecture underlying parental behaviour.

SummaryParenting is essential for the survival and wellbeing of mammalian offspring but we lack a circuit-level understanding of how distinct components of this behaviour are orchestrated. Here we investigate how Galanin-expressing neurons in the medial preoptic area (MPOAGal) coordinate motor, motivational, hormonal and social aspects of parenting. These neurons integrate inputs from a large number of brain areas, whose activation depends on the animal’s sex and reproductive state. Subsets of MPOAGal neurons form discrete pools defined by their projection sites. While the MPOAGal population is active during all episodes of parental behaviour, individual pools are tuned to characteristic aspects of parenting. Optogenetic manipulation of MPOAGal projections mirrors this specificity, affecting discrete parenting components. This functional organization, reminiscent of the control of motor sequences by pools of spinal cord neurons, provides a new model for how discrete elements of a social behaviour are generated at the circuit level.

Decomposer food web in a deciduous forest shows high share of generalist microorganisms and importance of microbial biomass recycling.

Forest soils represent important terrestrial carbon (C) pools where C is primarily fixed in the plant-derived biomass but it flows further through the biomass of fungi and bacteria before it is lost from the ecosystem as CO2 or immobilized in recalcitrant organic matter. Microorganisms are the main drivers of C flow in forests and play critical roles in the C balance through the decomposition of dead biomass of different origins. Here, we track the path of C that enters forest soil by following respiration, microbial biomass production, and C accumulation by individual microbial taxa in soil microcosms upon the addition of 13C-labeled biomass of plant, fungal, and bacterial origin. We demonstrate that both fungi and bacteria are involved in the assimilation and mineralization of C from the major complex sources existing in soil. Decomposer fungi are, however, better suited to utilize plant biomass compounds, whereas the ability to utilize fungal and bacterial biomass is more frequent among bacteria. Due to the ability of microorganisms to recycle microbial biomass, we suggest that the decomposer food web in forest soil displays a network structure with loops between and within individual pools. These results question the present paradigms describing food webs as hierarchical structures with unidirectional flow of C and assumptions about the dominance of fungi in the decomposition of complex organic matter.

Muscle and liver transcriptome characterization and genetic marker discovery in the farmed meagre, Argyrosomus regius

Meagre (Argyrosomus regius), a teleost fish of the family Sciaenidae, is part of a group of marine fish species considered new for Mediterranean aquaculture representing the larger fish cultured in the region. Meagre aquaculture started ~25years ago in West Mediterranean, and the supply of juveniles has been dominated by few hatcheries. This fact has raised concerns on possible inbreeding, urging the need for genetic information on the species and for an assessment of the polymorphisms found in the genome. To that end we characterized the muscle and liver transcriptome of a pool of meagre individuals, from different families and phenotypic size, to obtain a backbone that can support future studies regarding physiology, immunology and genetics of the species. The assembled transcripts were assigned to a wide range of biological processes including growth, reproduction, metabolism, development, stress and behavior. Then, to infer its genetic diversity and provide a catalogue of markers for future use, we scanned the reconstructed transcripts for polymorphic genetic markers. Our search revealed a total of 42,933 high quality SNP and 20,581 STR markers. We found a relatively low rate of polymorphism in the transcriptome that may indicate that inbreeding has taken place. This study has led to a catalogue of genetic markers at the expressed part of the genome and has set the ground for understanding growth and other traits of interest in meagre.

New rapid one-step PCR diagnostic assay for Plasmodium falciparum infective mosquitoes

An essential component of malaria vector control programmes is the detection of Plasmodium falciparum within its mosquito vectors, particularly in the salivary glands where the infective sporozoites reside. Several protocols have been developed for this purpose; however they require dissection of mosquito specimens prior to analysis. Here, a novel one-step RT-qPCR TaqMan diagnostic assay was developed for mosquitoes with infective Plasmodium falciparum sporozoites in the salivary glands. It is based on detection of the sporozoite-specific Pfslarp and Pfplp1 gene transcripts. These transcripts were chosen based on bioinformatics analysis, and experimentally verified to be overexpressed in the salivary gland sporozoite stage of the parasite compared to other mosquito parasite stages. The proof of principle and the performance of the assay were demonstrated using RNAlater preserved mosquito samples. Tests of analytical sensitivity showed the novel TaqMan assay to be 100% accurate, although its performance in the field needs to be further demonstrated. This method has no requirement for dissection and post-PCR processing and thus is simple and rapid to perform in individual mosquitoes or mosquito pools. It can be used in single or multiplex formats also targeting additional markers expressed in different tissues, such as detoxification enzymes associated with insecticide resistance.

Nutrient Stress Small-Molecule Screening Platform for Escherichia coli.

Clinically approved antibiotics target a narrow spectrum of cellular processes, namely cell wall synthesis, DNA replication, and protein synthesis. Numerous screens have been designed to identify inhibitors that target one of these cellular processes. Indeed, this narrow range of drug mechanisms and a reliance on chemical classes discovered many decades ago are thought to be principally responsible for the current crisis of antibiotic drug resistance. Seeking to expand the target base of antibacterial drug discovery, we developed a nutrient stress screening platform that identifies inhibitors of the growth of in Escherichia coli under nutrient limitation. Under nutrient stress, bacteria require an expanded biosynthetic capacity that includes the synthesis of amino acids, vitamins, and nucleobases. Growing evidence suggests that these processes may be indispensable to certain pathogens and at particular sites of infection. Indeed, more than 100 biosynthetic enzymes become indispensable to E. coli grown under nutrient stress in vitro. The screening platform described here puts a focus on these novel targets for new antibiotics and prioritizes growth inhibitory compounds that can be suppressed by individual nutrients and pools thereof.

,,,Equity in Contemporary Immigration Enforcement: Defining Contributions and Countering Criminalization

Since the 2016 Presidential election, discussions of immigration policy and enforcement have taken center stage in the public debate. In contrast to the Obama administration, which had articulated specific priorities for removal, the Trump administration has significantly expanded its enforcement targets. Indeed, high-level officials have confirmed that virtually anyone who is in the country without authorization is susceptible to removal. To make its case for enhanced immigration enforcement, the current administration has deployed familiar tropes regarding immigrant criminality and dangerousness. This rhetoric, operationalized through varied structures of criminalization, has shrunk the pool of individuals who can argue against removal, notwithstanding contributions or connections to the United States. The current political moment invites deeper reflection about the equities that should insulate undocumented noncitizens from removal, and how those equities should be assessed vis-a-vis allegations of criminality. Over the years, scholars have articulated a range of factors that justify protection from removal or even regularization of status. This article builds upon that literature, and presents a more nuanced typology of the societal contributions that should weigh against removal, along with the theories that undergird noncitizens’ claims in this context. A second purpose of this article is to expose how the powerful trend of criminalization has begun to infect even the limited space where discretion can be exercised, and has converted seemingly favorable conduct into undesirable criminal activity. This development, which is examined through the lens of several case studies, enables exclusion and expulsion, and disrupts the incentives that noncitizens have to contribute to and support the state.