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Invasive Non-Native Species (INNS) can co-transport externally and internally other organisms including viruses, bacteria and other eukaryotes (including metazoan parasites), collectively referred to as the symbiome. These symbiotic organisms include pathogens, a small minority of which are subject to surveillance and regulatory control, but most of which are currently unscrutinised and/or unknown. The (putatively) pathogenic symbionts co-transported by an INNS host may be latent or associated with asymptomatic infection and unable to cause disease in the INNS, but may be opportunistic pathogens of other hosts, causing impact to one or more hosts in their new range. These pathogens potentially pose diverse risks to other species, with implications for increased epidemiological risk to agriculture and aquaculture, wildlife/ecosystems, and human health (zoonotic diseases). Aquatic INNS and their symbionts have many introduction pathways, including commodity and trade (releases, escapes, contaminant), transport (stowaway), and dispersal (corridor, unaided). The risks and impacts arising from co-transported pathogens, including other symbionts of unknown pathogenic virulence, remain largely unexplored, unlegislated, and difficult to identify and quantify. Here, we propose a workflow to determine any known and potential pathogens of aquatic INNS. This workflow acts as a prerequisite for assessing the nature and risk posed by co-transported symbionts of INNS. A better understanding of co-transported organisms, the risks they pose and their impact, is necessary to inform policy and INNS risk assessments. This leap in evidence will be instrumental to devise an appropriate set of statutory responsibilities with respect to these symbionts, and to underpin new and more effective legislative processes relating to the disease screening and risk assessment of INNS.

BackgroundAltered resting state functional connectivity (FC) involving the anterior insula (aINS), a key node in the salience network, has been reported consistently in autism.MethodHere we examined, for the first time, FC between the aINS and the whole brain in a sample of full-term, postmenstrual age (PMA) matched neonates (mean 44.0 weeks, SD=1.5) who due to family history have high likelihood (HL) for developing autism (n=12) and in controls (n=41) without family history of autism (low likelihood, LL). Behaviors associated with autism were evaluated between 12 and 18 months (M=17.3 months, SD=2.5) in a subsample (25/53) of participants using the First Year Inventory (FYI).ResultsCompared to LL controls, HL neonates showed hypoconnectivity between left aINS and left amygdala. Lower connectivity between the two nodes was associated with higher FYI risk scores in the social domain (r(25) = −.561, p=.003) and this association remained robust when maternal mental health factors were considered. Considering that a subsample of LL participants (n=14/41) underwent brain imaging during the fetal period at PMA 31 and 34 weeks, in an exploratory analysis, we evaluated prospectively development of the LaINS-Lamy connectivity and found that the two areas strongly coactivate throughout the third trimester of pregnancy.ConclusionsThe study identifies left lateralized anterior insula - amygdala connectivity as a potential target of further investigation into neural circuitry that enhances likelihood of future onset of social behaviors associated with autism during neonatal and potentially prenatal periods.

This study provides a morphological, ultrastructural, and phylogenetic characterization of a novel micro-eukaryotic parasite (2.3–2.6 µm) infecting amphipod genera Echinogammarus and Orchestia. Longitudinal studies across two years revealed that infection prevalence peaked in late April and May, reaching 64% in Echinogammarus sp. and 15% in Orchestia sp., but was seldom detected during the rest of the year. The parasite infected predominantly hemolymph, connective tissue, tegument, and gonad, although hepatopancreas and nervous tissue were affected in heavier infections, eliciting melanization and granuloma formation. Cell division occurred inside walled parasitic cysts, often within host hemocytes, resulting in hemolymph congestion. Small subunit (18S) rRNA gene phylogenies including related environmental sequences placed the novel parasite as a highly divergent lineage within Class Filasterea, which together with Choanoflagellatea represent the closest protistan relatives of Metazoa. We describe the new parasite as Txikispora philomaios n. sp. n. g., the first confirmed parasitic filasterean lineage, which otherwise comprises four free-living flagellates and a rarely observed endosymbiont of snails. Lineage-specific PCR probing of other hosts and surrounding environments only detected T. philomaios in the platyhelminth Procerodes sp. We expand the known diversity of Filasterea by targeted searches of metagenomic datasets, resulting in 13 previously unknown lineages from environmental samples., Ministerio de Economía y Competitividad, Grant/Award Number: BFU2017- 90114- P; Horizon 2020 Framework Programme, Grant/Award Number: 747789; Department for Environment, Food and Rural Affairs, UK Government, Grant/Award Number: FB002A and FC1214; Hezkuntza, Hizkuntza Politika Eta Kultura Saila, Eusko Jaurlaritza, Grant/Award Number: PRE_2016_2_0124; Agencia Estatal de Investigación (AEI); Fondo Europeo de Desarrollo Regional (FEDER); Ayuda Juan de la Cierva—Incorporación, Grant/Award Number: IJC2018- 036657- I

Examination of faecal material has demonstrated how a broad range of organisms are distributed by bird movements. Such research has largely focused on dispersal of plant seeds by frugivores and of freshwater organisms by waterbirds. However, with few exceptions (e.g. avian influenza, Ebola virus), there is a dearth of evidence for transport of parasites and pathogens. High-throughput sequencing methods now provide a powerful means of addressing this knowledge gap by elucidating faecal contents in unprecedented detail. We collected faeces excreted by a range of migratory waterbirds in south-west Spain and pooled faecal DNA to create libraries reflective of feeding behavior. We created sets of libraries using high-throughput metagenomic and amplicon sequencing. For the latter we employed two sets of primers to broadly target the V4 region of the 18S rRNA gene (one set amplifying the region across all eukaryotes, the other excluding amplification of metazoans). Libraries revealed a wide diversity of eukaryotes, including parasites of the faecal producers themselves, parasites of food items, or those incidentally ingested. We also detected novel microbial eukaryotic taxa and found that parasite assemblage profiles were relatively distinct. Comparing the performance of the methods used supports their joint use for future studies of diversity and abundance. Because viable stages of many parasites are likely to be present in faeces, our results suggest significant levels of bird-mediated dispersal of parasites (both from avian and other hosts). Our methods revealed much hidden biodiversity, and allowed identification of the individuals who produced the faecal samples to species level, facilitating the study of interaction networks.

Contact tracing in the UK for Covid-19 is performed by NHS Test and Trace (NHSTT) via telephone or email. This study estimates how many patients who have been admitted to hospital are not reached by NHSTT and the number of their contacts who were not advised to self-isolate.Medical Student volunteers conducted face to face interviews with patients diagnosed with Covid-19 on an infectious diseases ward. Data on their close contacts were sent to NHSTT.20 cases were enrolled. 13(65%) did not engage with NHSTT, 4(20%) because they had no positive PCR, 9(45%) because of severity of illness, language or intellectual difficulties. 49 close contacts were identified of whom 33(67%) were from cases who had not engaged with NHSTT. “Backwards” contacts tracing information was collected from 11(55%) cases and 8(40%) gave detailed information.These data suggest that NHSTT fails to engage nearly two thirds of Covid-19 in-patients and fails to advise two thirds of their close contacts to self isolate.Volunteers used face to face interviews to overcome false negative tests, illness and communication problems to identify both close contacts and data on sources of infection.

This study provides a morphological, ultrastructural, and phylogenetic characterization of a novel micro-eukaryotic parasite (2.3-2.6 µm) infecting generaEchinogammarusandOrchestia. Longitudinal studies across two years revealed that infection prevalence peaked in late April and May, reaching 64% inEchinogammarussp. and 15% inOrchestiasp., but was seldom detected during the rest of the year. The parasite infected predominantly haemolymph, connective tissue, tegument, and gonad, although hepatopancreas and nervous tissue were affected in heavier infections, eliciting melanization and granuloma formation. Cell division occurred inside walled parasitic cysts, often within host haemocytes, resulting in haemolymph congestion. Small subunit (18S) rRNA gene phylogenies including related environmental sequences placed the novel parasite as a highly divergent lineage within Class Filasterea, which together with Choanoflagellatea represent the closest protistan relatives of Metazoa. We describe the new parasite asTxikispora philomaiosn. sp. n. g., the first confirmed parasitic filasterean lineage, which otherwise comprises four free-living flagellates and a rarely observed endosymbiont of snails. Lineage-specific PCR probing of other hosts and surrounding environments only detectedT. philomaiosin the platyhelminthProcerodessp. We expand the known diversity of Filasterea by targeted searches of metagenomic datasets, resulting in 13 previously unknown lineages from environmental samples.

High-throughput DNA metabarcoding of amplicon sizes below 500 bp has revolutionized the analysis of environmental microbial diversity. However, these short regions contain limited phylogenetic signal, which makes it impractical to use environmental DNA in full phylogenetic inferences. This lesser phylogenetic resolution of short amplicons may be overcome by new long-read sequencing technologies. To test this idea, we amplified soil DNA and used PacBio Circular Consensus Sequencing (CCS) to obtain an ~4500-bp region spanning most of the eukaryotic small subunit (18S) and large subunit (28S) ribosomal DNA genes. We first treated the CCS reads with a novel curation workflow, generating 650 high-quality operational taxonomic units (OTUs) containing the physically linked 18S and 28S regions. To assign taxonomy to these OTUs, we developed a phylogeny-aware approach based on the 18S region that showed greater accuracy and sensitivity than similarity-based methods. The taxonomically annotated OTUs were then combined with available 18S and 28S reference sequences to infer a well-resolved phylogeny spanning all major groups of eukaryotes, allowing us to accurately derive the evolutionary origin of environmental diversity. A total of 1,019 sequences were included, of which a majority (58%) corresponded to the new long environmental OTUs. The long reads also allowed us to directly investigate the relationships among environmental sequences themselves, which represents a key advantage over the placement of short reads on a reference phylogeny. Together, our results show that long amplicons can be treated in a full phylogenetic framework to provide greater taxonomic resolution and a robust evolutionary perspective to environmental DNA.

Therapeutic mammaplasty (TM) may be an alternative to mastectomy, but few well designed studies have evaluated the success of this approach or compared the short-term outcomes of TM with mastectomy with or without immediate breast reconstruction (IBR). Data from the national iBRA-2 and TeaM studies were combined to compare the safety and short-term outcomes of TM and mastectomy with or without IBR.The subgroup of patients in the TeaM study who underwent TM to avoid mastectomy were identified, and data on demographics, complications, oncology and adjuvant treatment were compared with those of patients undergoing mastectomy with or without IBR in the iBRA-2 study. The primary outcome was the percentage of successful breast-conserving procedures in the TM group. Secondary outcomes included postoperative complications and time to adjuvant therapy.A total of 2916 patients (TM 376; mastectomy 1532; mastectomy and IBR 1008) were included in the analysis. Patients undergoing TM were more likely to be obese and to have undergone bilateral surgery than those having IBR. However, patients undergoing mastectomy with or without IBR were more likely to experience complications than the TM group (TM: 79, 21·0 per cent; mastectomy: 570, 37·2 per cent; mastectomy and IBR: 359, 35·6 per cent; P 0·001). Breast conservation was possible in 87·0 per cent of patients who had TM, and TM did not delay adjuvant treatment.TM may allow high-risk patients who would not be candidates for IBR to avoid mastectomy safely. Further work is needed to explore the comparative patient-reported and cosmetic outcomes of the different approaches, and to establish long-term oncological safety.La mamoplastia terapéutica (therapeutic mammaplasty, TM) puede ser una alternativa a la mastectomía, pero hay pocos estudios bien diseñados que hayan evaluado el éxito de esta estrategia o hayan comparado los resultados a corto plazo de la TM con la mastectomía con o sin (+/-) reconstrucción mamaria inmediata (immediate breast reconstruction, IBR). Para comparar la seguridad y los resultados a corto plazo de la TM y la mastectomía +/- IBR se combinaron los datos de los estudios nacionales iBRA-2 y TeaM. MÉTODOS: En el estudio TeaM se identificó el subgrupo de pacientes al que se realizó una TM para evitar la mastectomía y se compararon los datos demográficos, las complicaciones, los resultados oncológicos y el tratamiento adyuvante con las pacientes sometidas a mastectomía +/- IBR del estudio iBRA-2. La variable principal fue el porcentaje de éxito de la cirugía conservadora de mama en el grupo TM. Las variables secundarias fueron las complicaciones postoperatorias y el intervalo de tiempo hasta el inicio del tratamiento adyuvante.Se incluyeron en el análisis 2.916 pacientes (TM n = 376; mastectomía n = 1.532; IBR n = 1.008). La TM era más frecuente en pacientes obesas o en las sometidas a cirugía bilateral en comparación con las pacientes con IBR. Sin embargo, las pacientes sometidas a una mastectomía +/- IBR tenían más probabilidades de desarrollar complicaciones que las del grupo TM (TM n = 79, 21,0%; mastectomía n = 570, 37,2%; mastectomía y IBR n = 359, 35,6%; P 0,001). La conservación de la mama fue posible en el 87% de las pacientes con TM y el procedimiento no retrasó el inicio del tratamiento adyuvante. CONCLUSIÓN: La TM puede permitir que pacientes de alto riesgo que no serían candidatas a IBR eviten la mastectomía de una forma segura. Se necesitan más trabajos para comparar los resultados percibidos por las pacientes y los estéticos de las diferentes estrategias terapéuticas y establecer la seguridad oncológica a largo plazo.

High-throughput environmental DNA metabarcoding has revolutionized the analysis of microbial diversity, but this approach is generally restricted to amplicon sizes below 500 base pairs. These short regions contain limited phylogenetic signal, which makes it impractical to use environmental DNA in full phylogenetic inferences. However, new long-read sequencing technologies such as the Pacific Biosciences platform may provide sufficiently large sequence lengths to overcome the poor phylogenetic resolution of short amplicons. To test this idea, we amplified soil DNA and used PacBio Circular Consensus Sequencing (CCS) to obtain a ~4500 bp region of the eukaryotic rDNA operon spanning most of the small (18S) and large subunit (28S) ribosomal RNA genes. The CCS reads were first treated with a novel curation workflow that generated 650 high-quality OTUs containing the physically linked 18S and 28S regions of the long amplicons. In order to assign taxonomy to these OTUs, we developed a phylogeny-aware approach based on the 18S region that showed greater accuracy and sensitivity than similarity-based and phylogenetic placement-based methods using shorter reads. The taxonomically-annotated OTUs were then combined with available 18S and 28S reference sequences to infer a well-resolved phylogeny spanning all major groups of eukaryotes, allowing to accurately derive the evolutionary origin of environmental diversity. A total of 1019 sequences were included, of which a majority (58%) corresponded to the new long environmental CCS reads. Comparisons to the 18S-only region of our amplicons revealed that the combined 18S-28S genes globally increased the phylogenetic resolution, recovering specific groupings otherwise missing. The long-reads also allowed to directly investigate the relationships among environmental sequences themselves, which represents a key advantage over the placement of short reads on a reference phylogeny. Altogether, our results show that long amplicons can be treated in a full phylogenetic framework to provide greater taxonomic resolution and a robust evolutionary perspective to environmental DNA.

Introduction There is a variety of testing methods described in the literature for the spinal accessory nerve (SAN). This study aims to evaluate side-to-side, gender, and BMI differences with surface recording from the upper and middle trapezius using a standard distance to the upper trapezius. Methods Subjects underwent bilateral SAN conduction testing with the active recording electrodes over the superior border of the upper trapezius, midway between the acromion and the C7 spinous process, and over the middle trapezius 3 cm medial to the vertebral border of the scapula. Results Mean latency values were 2.17 ± 0.22 msec and 3.14 ± 0.40 msec for the upper and middle trapezius, respectively. Mean amplitude values were 8.02 ± 2.2 mV for the upper trapezius and 3.96 ± 1.77 mV for the middle trapezius. The mean side-to-side latency difference was 7.8% for the upper and 9.5% for the middle trapezius, while the mean side-to-side amplitude difference was 18.2% for the upper and 37.6% for the middle trapezius. BMI had a significant inverse effect on upper and middle trapezius amplitudes such that both males and females with lower BMI had larger amplitudes. There was a significant gender difference for upper and middle trapezius latency with faster latency values observed in females. Conclusions SAN conduction with surface recording from the upper and middle trapezius is well tolerated. Side-to-side differences may be the best way to evaluate both amplitude and latency, so bilateral testing is essential in light of anatomical variation and BMI effects on amplitude.

Background: Owing to this improved technology and peri-implantation care around 90% of patients being considered for mechanical circulatory support receive an LVAD. Although LVAD support is associated with improved survival and quality of life, infectious complications remain a major limitation. The exact incidence of LVAD-related and non-VAD related infections among patients with VADs is unknown. Several potential factors make patients with VADs more prone to infectious complications, most obvious being the presence of an implanted device. Temporary alterations in the host immune system, including changes in T-cell function and quantity, have been observed after VAD implantation and may contribute to development of VAD-related infections. The optimal approach to empiric antibiotic therapy and duration of treatment has yet to be fully elucidated. Current treatment recommendations are based on observational studies and expert opinion, as no randomized controlled trials have evaluated empiric treatment of VAD infections. In the past several years we have expanded our understanding of driveline infections in VAD patients. However, information about non-drive line infections in VAD patients remains limited. Study Objectives: This is a quality-improvement (QI) project to look for preventable causes of driveline as well as non-driveline infections (i.e. VAD related and non-VAD related infections) among LVAD patients cared for at University of Kansas Medical Center. Primary Objectives a) Investigate the incidence and types of non-driveline infections among LVAD patients within the first six months of their VAD implantation. b) Investigate the incidence of driveline infections among LVAD patients within the first six months of their VAD implantation. c) Identify potentially preventable causes of driveline and non-driveline infections and propose interventions to reduce the future incidence of such infections. Secondary Objective a) Share our findings with our leadership with the ultimate goal of developing recommendations on empiric antibiotic treatments based on patient-specific risk factors Study Methods: We will perform retrospective chart review of LVAD patients to establish the incidence and types of infections among these patients within the first six months of LVAD implantation. Study will only include patients who received their LVAD implantations at University of Kansas Medical Center between October 2015 and October 2017. Data collection will include patient demographics, co-morbidities and risk factors for infections, as well as infection-focused information including site of acquisition and time to onset, causative organism, and microbiological data (e.g., cultures and sensitivities, viral panels, MIC, MRSA screen) and duration of therapy.

A 48-year-old woman presented to the Accident and Emergency department with a 4 month history of headaches, nausea and dizziness. She was found to have severe hypertension and hypokalaemia. Extensive investigations did not find any secondary cause for hypertension. The patient was discharged with oral doxazosin therapy which controlled the blood pressure. Before the follow-up appointment at the hypertension clinic, the patient and her husband identified that her headaches coincided with liquorice tea consumption of up to three cups per day. This information was not obtained in the clinical assessment. The patient is now headache and medication free after cessation of liquorice tea. Liquorice ingestion is often a forgotten reversible cause of hypertension. A good history is key to this diagnosis.

Do reach-to-grasp (prehension) movements require a metric representation of three-dimensional (3D) layouts and objects? We propose a model relying only on direct sensory information to account for the planning and execution of prehension movements in the absence of haptic feedback and when the hand is not visible. In the present investigation, we isolate relative motion and binocular disparity information from other depth cues and we study their efficacy for reach-to-grasp movements and visual judgments. We show that (i) the amplitude of the grasp increases when relative motion is added to binocular disparity information, even if depth from disparity information is already veridical, and (ii) similar distortions of derived depth are found for haptic tasks and perceptual judgments. With a quantitative test, we demonstrate that our results are consistent with the Intrinsic Constraint model and do not require 3D metric inferences (Domini, Caudek, & Tassinari, 2006). By contrast, the linear cue integration model (Landy, Maloney, Johnston, & Young, 1995) cannot explain the present results, even if the flatness cues are taken into account.

One frequently reported result is that, for perceptual tasks, the amount of perceived depth is larger when motion and disparity cues are presented together than when only one of these depth cues is shown at a time. It is not known whether this increase of perceived depth in the combined condition is also found when depth estimation is aided by action. The action of reaching-to-grasp might provide additional information about the depth of target objects, thus further disambiguating the estimation of 3D depth from projected images. We tested this through an experiment investigating the problem of integration of motion and stereoscopic depth information for two manual depth estimation tasks: one based in perception [depth estimated through fingers adjustments], and one in action [depth estimated through reach-to-grasp]. In both tasks, observers viewed a display composed of three vertical lines embedded in a cloud of random dots; two flanking lines were positioned at fixation distance with the third line midway between the two located in depth (in front of the flankers). Observers estimated five different levels of depth separation between the flankers and the central line. 3D information was provided either by binocular disparities (stereo only), image velocities (motion only) or both (stereo-motion). Haptic feedback was provided in one fourth of the trials to calibrate depth estimates and ensure that normal visual-open-loop prehension occurred even in trials lacking haptic feedback (Bingham, Coats, Mon-Williams 2007). The results show that in the combined condition, observers perceived a larger amount of estimated depth with both the perception and action tasks.

Daphnia provides an easily visible heart which responds rapidly to soluble drugs. Stroboscopic illumination of the animal enables students to make accurate heart rate counts

After transferring leaf, pith, and stemcortex tissues ofNicotiana tabacum L. cv. "Havana 425" from a complete medium containing auxin and cytokinin to an inductive medium with auxin deleted, there is lag phase of approx. 14d followed by a linear phase in which the rate of bud initiation is constant. The incidence of buds formed is very low, approx. one bud per 10(3) or 10(4) cells. Statistical analysis of the distribution of buds among explants and subcloning experiments provide evidence that the paucity of buds results from neither negative interactions among bud forming centers nor a paucity of cells with the potential for organogenesis. Our results are consistent with the hypothesis that the frequency of bud initiation is determined by the availability of competent cells, by position effects, or by a combination of both mechanisms.

Rhizarian ‘Novel Clade 10’ (NC10) is frequently detected by 18S rRNA gene sequencing studies in freshwater planktonic samples. We describe a new genus and two species of eukaryovorous biflagellate protists, Aquavolon hoantrani n. gen. n. sp. and A. dientrani n. gen. n. sp., which represent the first morphologically characterized members of NC10, here named Aquavolonida ord. nov. The slightly metabolic cells possess naked heterodynamic flagella, whose kinetosomes lie at a right angle to each other and are connected by at least one fibril. Unlike their closest known relative Tremula longifila, they rotate around their longitudinal axis when swimming and only very rarely glide on surfaces. Screening of a wide range of environmental DNA extractions with lineage‐specific PCR primers reveals that Aquavolonida consists of a large radiation of protists, which are most diversified in freshwater planktonic habitats and as yet undetected in marine environments. Earlier‐branching lineages in Aquavolonida include less frequently detected organisms from soils and freshwater sediments. The 18S rRNA gene phylogeny suggests that Aquavolonida forms a common evolutionary lineage with tremulids and uncharacterized ‘Novel Clade 12’, which likely represents one of the deepest lineages in the Rhizaria, separate from Cercozoa (Filosa), Endomyxa, and Retaria.