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This study was designed to identify abnormalities in the electroencephalograms (EEGs) recorded from stranded California sea lions (Zalophus californianus) with suspected domoic acid (DA) toxicosis. Recordings from animals presenting for non-neurological issues were also obtained to better understand the normal EEG (background activity and transient events) in this species, as, to date, studies have focused on examining natural sleep in pinnipeds. Most animals were sedated for electrode placement and EEG acquisition with some receiving antiepileptic medications or isoflurane during the procedure. A total of 103 recordings were read and scored from 0 (normal) to 3 (severely abnormal). Epileptiform discharges, consisting of spikes, sharp waves, slow waves, and/or spike waves, were present in all EEGs with scores of 1, 2, or 3. The distribution of these events over the scalp varied. While often generalized, others were lateralized over one hemisphere, bifrontal, bioccipital, and/or bitemporal, while some discharges were multifocal. Findings were different between sea lions and occasionally changed within the EEG on a given sea lion. No clinical seizures were observed during the recording but a few sea lions had findings consistent with electroencephalographic seizures. When available, supporting diagnostic results obtained from magnetic resonance imaging (MRI) and/or necropsy/histopathology were described, as well as the status of those sea lions that recovered and were released with satellite tags.

Calanoid copepods comprise around 90% of Arctic zooplankton biomass and are fundamental to the ecological and biogeochemical functioning of high-latitude pelagic ecosystems. They accumulate lipid reserves during the productive months and represent an energy-rich food source for higher trophic levels. Rapidly changing climate in the Arctic may alter the quantity and composition of the food environment for one of the key copepod species, Calanus finmarchicus, with as yet unquantified effects on its production. Here we present rates of feeding and egg production in female C. finmarchicus exposed to the range of feeding conditions encountered across the Fram Strait in May/June 2018. Carbon (C) budgets were constructed and used to examine the relationship between feeding and growth (= egg production) in these animals. C-specific ingestion rates (mean ± standard deviation) were highly variable, ranging from 0.015 ± 0.004 to 0.645 ± 0.017 day-1 (mean = 0.295 ± 0.223 day-1), and were positively correlated with food availability. C-specific egg production rates ranged from 0.00 to 0.049 day-1 (mean = 0.012 ± 0.011) and were not correlated with either food availability or ingestion rate. Calculated gross growth efficiencies (GGE: growth/ingestion) were low, 0.12 ± 0.13 (range = 0.01 to 0.39). The assembled C budgets indicate that the average fraction of ingested food that was surplus to the requirements for egg production, respiration and losses to faecal pellets was 0.17 ± 0.42. We suggest that this excess occurred, at least in part, because many of the incubated females were still undergoing the energetically (C-) expensive process of gonad maturation at the time of sampling, an assertion that is supported by the relatively high C:N (nitrogen) ratios of the incubated females, the typically low egg production rates, and gonad maturation status. Ontogenetic development may thus explain the large variability seen in the relationship between egg production and ingestion. The apparently excessive ingestion rates may additionally indicate that recently moulted females must acquire additional N via ingestion to complete the maturation process and begin spawning. Our results highlight the need for improved fundamental understanding of the physiology of high-latitude copepods and its response to environmental change.

To understand the sources and migration pattern of PM2.5 in the Yangtze River Delta (YRD), China, the total carbon (TC) and total nitrogen (TN) concentrations and the corresponding stable isotope ratios (δ13CTC and δ15NTN) were determined in aerosol samples simultaneously collected from August 2014 to April 2015 at three different locations (Shanghai, Ningbo, Nanjing). Ningbo and Shanghai are geographically closer, the research results precisely divide Nanjing and the other two cities into two categories. Nanjing has a higher proportion of nitrogen in PM2.5 (13.2–15.3%) than Shanghai and Ningbo (8.6–12.6%), and the correlation analysis shows that nitrogen components (mainly ammonium nitrogen) might be the main driving force for the formation of PM2.5. The isotopes were proven to be sensitive sensors to reflect the impact of special events on PM2.5. For example, compared to other seasons, δ13CTC in autumn in the three cities are relatively depleted, indicating an input from biomass combustion to PM2.5 at this time. On New Year’s Eve, three cities simultaneously observed enriched δ13CTC due to the burning of fireworks. During the Qingming Festival, abnormally depleted nitrogen isotope ratios were observed, reflecting the vehicle exhaust pollution caused by people’s short travel. Isotopes are also used to trace the transport process of PM2.5. Postponing the sampling date in Nanjing by one day increased the linear fit (r2) of δ13CTC between Nanjing and Ningbo from 0.03 to 0.75, while that of δ15NTN improved from 0.16 to 0.63, which means PM2.5 might transport from Nanjing to Shanghai and Ningbo, and the transfer time takes one day.

Lipid-storing copepods are fundamental to the functioning of marine ecosystems, transferring energy from primary producers to higher trophic levels and sequestering atmospheric carbon (C) in the deep ocean. Quantifying trophic transfer and biogeochemical cycling by copepods requires improved understanding of copepod metabolic rates in both surface waters and during lipid-fueled metabolism over winter. Here we present new biomass turnover rates of C and nitrogen (N) in Calanoides acutus, Calanoides natalis, Calanus glacialis and Calanus hyperboreus alongside published data for Calanus finmarchicus and Calanus pacificus. Turnover rates in metabolically active animals, normalised to 10°C, ranged between 0.007 – 0.105 d-1 and 0.004 – 0.065 d-1 for C and N, respectively. Turnover rates of C were typically faster than those for N, supporting the understanding that non-protein C, e.g. lipid, is catabolised faster than protein. Re-analysis of published data indicates that inactive, overwintering C. finmarchicus turn over wax ester lipids at a rate of 0.0016 d-1. These and other basal rate data will facilitate the mechanistic representation of copepod physiology in global biogeochemical models, thereby reducing uncertainties in our predictions of future ocean ecosystem functioning and C sequestration.

YAP1 gene fusions are found in subgroups of paediatric ependymomas. Here the authors show that YAP1 activation in NeuroD6 positive neuronal precursor cells can induce ependymoma-like tumours in mice.

Western Central European Loess-Palaeosol-Sequences (LPS) provide valuable terrestrial records of palaeoenvironmental conditions, which formed in response to variability in the North Atlantic climate systems. Over the last full glacial cycle (∼130 ka), climate oscillations within these systems are best documented in deep sea- and ice cores; the responses of terrestrial systems are not yet fully understood. A better understanding of metabolism governing input and output variables of organic- and inorganic C pools is, however, crucial for investigating landscape-atmospheric feedback processes and in particularly, for understanding the formation of calcareous LPS as environmental archives. Here we quantify the contributions of primary carbonates (PC) and secondary carbonates (SC) to the overall inorganic carbon pool down a LPS at the Schwalbenberg site, based on the natural abundance ratio of stable carbon isotopes (δ13C) and contents of the organic- and inorganic C pools. This facilitates detailed insights into the carbonate metabolism and hence, loessification and percolation processes. PC accumulate predominantly in cold phases during periods of reduced biological activity and become leached during wetter and warmer periods contemporary with higher rates of SC re-precipitation and total organic carbon (TOC) increases due to enhanced biomass production. We find that mineral dust input is most significant during stadials, as well as toward the end of warmer interstadials, characterised by gradual cooling back to stadial conditions. Pedogenesis in the Schwalbenberg LPS kept pace with surface accumulation of mineral dust. This indicates that palaeosols are of accretionary nature, which gives raise to the idea of incorporation of former topsoils in preserved subsoil horizons. Our study decodes fundamental aspects of the link between atmospheric dust circulation and terrestrial records in western Central Europe. In addition, interdependencies between factors governing the regional moisture budged and LPS can be reconstructed in a more holistic way than before.

Abstract The expansion of global aquaculture activities is important for the wellbeing of future generations in terms of employment and food security. Rearing animals in open-exchange cages permits the release of organic wastes, some of which ultimately reaches the underlying sediments. The development of rapid, quantitative and objective monitoring techniques is therefore central to the environmentally sustainable growth of the aquaculture industry. Here, we demonstrate that fish farm-derived organic wastes can be readily detected at the seafloor by quantifying sediment phospholipid fatty acids (PLFAs) and their carbon stable isotope signatures. Observations across five farms reveal that farm size and/or distance away from it influence the spatial distribution of the generated organic wastes and their effect on benthic bacterial biomass. Comparison to the isotopic signatures of fish feed-derived PLFAs indicates that 16:0 and 18:1(n-9) are potential biomarkers for fish farm-derived organic wastes. Our results suggest that stable isotope analysis of sediment PLFAs has potential for monitoring the environmental performance of aquaculture activities, particularly given the increasing prevalence of terrigenous organic matter in aquaculture feed stocks because it is isotopically district to marine organic matter.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Damage-associated molecular patterns (DAMPs) are molecules exposed or released by dead cells that trigger or modulate immunity and tissue repair. In vertebrates, the cytoskeletal component F-actin is a DAMP specifically recognised by DNGR-1, an innate immune receptor. Previously we suggested that actin is also a DAMP in Drosophila melanogaster by inducing STAT-dependent genes (Srinivasan et al., 2016). Here, we revise that conclusion and report that α-actinin is far more potent than actin at inducing the same STAT response and can be found in trace amounts in actin preparations. Recombinant expression of actin or α-actinin in bacteria demonstrated that only α-actinin could drive the expression of STAT target genes in Drosophila. The response to injected α-actinin required the same signalling cascade that we had identified in our previous work using actin preparations. Taken together, these data indicate that α-actinin rather than actin drives STAT activation when injected into Drosophila.

Squamous cell carcinoma (SCC) can progress to malignant metastatic cancer, including an aggressive subtype known as spindle cell carcinoma (spSCC). spSCC formation involves epithelial-to-mesenchymal transition (EMT), yet the molecular basis of this event remains unknown. The transcriptional co-activator YAP undergoes recurrent amplification in human SCC and overexpression of YAP drives SCC formation in mice. Here, we show that human spSCC tumours also feature strong nuclear localisation of YAP and overexpression of activated YAP (NLS-YAP-5SA) with Keratin-5 (K5-CreERt) is sufficient to induce rapid formation of both SCC and spSCC in mice. spSCC tumours arise at sites of epithelial scratch wounding, where tumour-initiating epithelial cells undergo EMT to generate spSCC. Expression of the EMT transcription factor ZEB1 arises upon wounding and is a defining characteristic of spSCC in mice and humans. Thus, the wound healing response synergises with YAP to drive metaplastic transformation of SCC to spSCC.

AimsBiological and lifestyle factors, such as daily rhythm, caffeine ingestion, recent infection, and antibiotic intake, have been shown to influence measurements of salivary cortisol (SC) and secretory immunoglobulin A (sIgA). Current methodology in unsynchronized, field-based biomarker studies does not take these effects into account. Moreover, very little is known about the combined effects of biological and lifestyle factors on SC and sIgA. This study supports development of a protocol for measuring biomarkers from saliva collected in field studies by examining the individual and combined effects of these factors on SC and sIgA.MethodAt three time points (start of the pre-season; start of playing season; and end of playing season), saliva samples were collected from the entire squad of 45 male players of an elite Australian Football club (mean age 22.8 ± 3.5 years). At each time, point daily rhythm and lifestyle factors were determined via a questionnaire, and concentrations of both SC and sIgA via an enzyme linked immuno-sorbent (ELISA) assay of saliva samples. In addition, player times to produce 0.5 mL of saliva were recorded.ResultsAnalysis of covariance of the data across the three time points showed that daily rhythm had a more consistent effect than the lifestyle factors of caffeine ingestion, recent infection, and antibiotic intake on SC, but not on sIgA. Data for sIgA and SC concentrations were then adjusted for the effects of daily rhythm and lifestyle factors, and correlational analysis of the pooled data was used to examine the relative effects of these two sources of influence on sIgA and SC. With the exception of time to produce saliva, the biological measures of stress were affected by players’ daily rhythms. When daily rhythm was taken into account the group of lifestyle factors did not have an additional effect.DiscussionIt is recommended that future studies measuring SC and sIgA make additional adjustments for the daily rhythm, in particular time since first sight of daylight, as small measurement errors of biomarkers can confound discrimination among study participants.

Damage-associated molecular patterns (DAMPs) are molecules released by dead cells that trigger sterile inflammation and, in vertebrates, adaptive immunity. Actin is a DAMP detected in mammals by the receptor, DNGR-1, expressed by dendritic cells (DCs). DNGR-1 is phosphorylated by Src-family kinases and recruits the tyrosine kinase Syk to promote DC cross-presentation of dead cell-associated antigens. Here we report that actin is also a DAMP in invertebrates that lack DCs and adaptive immunity. Administration of actin to Drosophila melanogaster triggers a response characterised by selective induction of STAT target genes in the fat body through the cytokine Upd3 and its JAK/STAT-coupled receptor, Domeless. Notably, this response requires signalling via Shark, the Drosophila orthologue of Syk, and Src42A, a Drosophila Src-family kinase, and is dependent on Nox activity. Thus, extracellular actin detection via a Src-family kinase-dependent cascade is an ancient means of detecting cell injury that precedes the evolution of adaptive immunity.

Dry mass production and persistence of Panicum maximum pastures depends on nitrogen supply. Defoliation influences N uptake and allocation patterns yet its effects on plasticity of N dynamics in P. maximum have not been investigated. Stable isotopes of N (15N) were used in order to test the hypothesis that defoliation in terms of proportion of the leaf area removed effects N mobilisation, uptake and allocation patterns in P. maximum. The plants were initially cut weekly to a height of either 0.15 m or 0.30 m for seven weeks. Eight weeks after the first defoliation, all plants were defoliated for a final time to remove 0, 25, 50, 75 or 100 % of the area of each individual leaf blade of the main tiller. Root N uptake was reduced when all leaf area was removed, but more lenient defoliation improved N uptake due to a positive effect on specific N uptake. Young leaves, side tillers and roots were the main sinks for N from root uptake. Roots of P. maximum became a net source of N for mobilisation immediately after severe defoliation. Root uptake was the main source of N for new growth in P. maximum plants. Allocation pattern of mobilised N was different from that of N derived from root uptake. It was concluded that adaptation of P. maximum to defoliation is related to plasticity of N uptake, mobilisation and allocation, but changes in N dynamics did not offset negative impacts of complete defoliation of the plants.

The recent April 2011 meeting of the G20 countries considered possible development of a global early warning system to avoid any future financial crisis. Psycho-economic factors are strong drivers of greed, fear and non-rational behavior and experience shows that they should not be excluded from such a project. Rational, logical behavior for attitude and actions has been an assumption in most financial models prior to the advent of the 2008 crisis. In recent years there has been an increasing interest in relating financial activity to phenomena in physics, turbulence, neurology and recent fMRI experiments show that cortical interactions for decisions are affected by previous experience. We use an extension of two Lotka-Volterra (LV) interactive equations used in a model for the 2008 crisis but now with fluctuation theory from chemical physics to interact the two previously used heterogenous interacting agents, the psycho-economic ratio CE of investor expectations (favourable/unfavourable) and the reality ratio of equity/debt. The model provides a variable, M, for uncertainties in CE arising from the ability of the economy to affect the financial sector. A condition obtained for keeping rates of change in M small to avoid divergence of spontaneous fluctuations, provides a quantifiable time dependent entity which can act as a warning of impending crisis. The conditional expression appears to be related to an extension of Ohm's law as in a recently discovered "chip" and memory; the memristor. The possible role of subthreshold legacies in CE from the previous crisis appears to be possible and related to recent neurological findings.

More than 50% of terrestrially-derived organic carbon (terrOC) flux from the continents to the ocean is remineralised in the coastal zone despite its perceived high refractivity. The efficient degradation of terrOC in the marine environment could be fuelled by labile marine-derived material, a phenomenon known as "priming effect", but experimental data to confirm this mechanism are lacking. We tested this hypothesis by treating coastal sediments with 13C-lignocellulose, as a proxy for terrOC, with and without addition of unlabelled diatom detritus that served as the priming inducer. The occurrence of priming was assessed by the difference in lignocellulose mineralisation between diatom-amended treatments and controls in aerobic sediment slurries. Priming of lignocellulose degradation was observed only at the initial stages of the experiment (day 7) and coincided with overall high microbial activity as exemplified by total CO2 production. Lignocellulose mineralisation did not differ consistently between diatom treatments and control for the remaining experimental time (days 14-28). Based on this pattern, we hypothesize that the faster initiation of lignocellulose mineralisation in diatom-amended treatments is attributed to the decomposition of accessible polysaccharide components within the lignocellulose complex by activated diatom degraders. The fact that diatom-degraders contributed to lignocellulose degradation was also supported by the different patterns in 13C-enrichment of phospholipid fatty acids between treatments. Although we did not observe differences between treatments in the total quantity of respired lignocellulose at the end of the experiment, differences in timing could be important in natural ecosystems where the amount of time that a certain compound is subject to aerobic degradation before burial to deeper anoxic sediments may be limited.

Copper is essential for healthy cellular functioning, but this heavy metal quickly becomes toxic when supply exceeds demand. Marine sediments receive widespread and increasing levels of copper contamination from antifouling paints owing to the 2008 global ban of organotin-based products. The toxicity of copper will increase in the coming years as seawater pH decreases and temperature increases. We used a factorial mesocosm experiment to investigate how increasing sediment copper concentrations and the presence of a cosmopolitan bioturbating amphipod, Corophium volutator, affected a range of ecosystem functions in a soft sediment microbial community. The effects of copper on benthic nutrient release, bacterial biomass, microbial community structure and the isotopic composition of individual microbial membrane [phospholipid] fatty acids (PLFAs) all differed in the presence of C. volutator. Our data consistently demonstrate that copper contamination of global waterways will have pervasive effects on the metabolic functioning of benthic communities that cannot be predicted from copper concentrations alone; impacts will depend upon the resident macrofauna and their capacity for bioturbation. This finding poses a major challenge for those attempting to manage the impacts of copper contamination on ecosystem services, e.g. carbon and nutrient cycling, across different habitats. Our work also highlights the paucity of information on the processes that result in isotopic fractionation in natural marine microbial communities. We conclude that the assimilative capacity of benthic microbes will become progressively impaired as copper concentrations increase. These effects will, to an extent, be mitigated by the presence of bioturbating animals and possibly other processes that increase the influx of oxygenated seawater into the sediments. Our findings support the move towards an ecosystem approach for environmental management.

The priming effect (PE) is a complex phenomenon which describes a modification (acceleration or retardation) in the mineralisation rate of refractory organic matter (OM) following inputs of labile material. PEs are well-studied in terrestrial ecosystems owing to their potential importance in the evolution of soil carbon stocks but have been largely ignored in aquatic systems despite the fact that the prerequisite for their occurrence, i.e. the co-existence of labile and refractory OM, is also true for sediments. We conducted stable isotope tracer experiments in continental margin sediments from the NE Atlantic (550-950 m) to study PE occurrence and intensity in relation to labile OM input. Sediment slurries were treated with increasing quantities of the (13)C-labelled diatom Thalassiosira rotula and PE was quantified after 7, 14 and 21 days. There was a stepwise effect of diatom quantity on its mineralisation although mineralisation efficiency dropped with increasing substrate amounts. The addition of diatomaceous OM yielded a negative PE (i.e. retardation of existing sediment OM mineralisation) at the end of the experiment regardless of diatom quantity. Negative PE is often the result of preferential utilisation of the newly deposited labile material by the microbial community ("preferential substrate utilization", PSU) which is usually observed at excessive substrate additions. The fact that PSU and the associated negative PE occurred even at low substrate levels in this study could be attributed to limited amounts of OM subject to priming in our study area (~0.2% organic carbon [OC]) which seems to be an exception among continental slopes (typically >0.5%OC). We postulate that PEs will normally be positive in continental slope sediments and that their intensity will be a direct function of sediment OC content. More experiments with varying supply of substrate targeting C-poor vs. C-rich sediments are needed to confirm these hypotheses.

Gas chromatography combustion isotope ratio mass spectrometry (GC-C-IRMS) was used to measure the (13)C/(12)C ratios of PLFAs at natural abundance levels from a temperate grassland nitrogen (N) and phosphorus (P) factorial fertilization experiment in northern Greece. In each plot two rhizosphere samples were derived centred around individual Agrostis capillaris and Prunella vulgaris plants. It was hypothesized that the isotopic signal of microbes that preferentially feed on recalcitrant litter such as fungi would be modified by fertilization more strongly than that of opportunistic microbes using labile C. Microbial community δ(13)C was affected by both P and N fertilization regime and plant species identity. However, we have been unable to detect significant nutrient effects on individual groups of microbes when analyzed separately in contrast to our original hypothesis. Intra-treatment variability, as evaluated from Hartley's F(max) tests in the five first PCA components axes as well as the size of the convex hulls in PCA scoreplots and Mahalanobis distances, was considerably higher in the non-fertilized controls. Moreover, a significant relationship was established between the change in PLFA abundances and their respective changes in δ(13)C for the aggregate of samples and those simultaneously fertilized with N and P. We conclude that use of compound specific isotope analysis in the absence of labelling represents a valuable and overlooked tool in obtaining an insight of microbial community functioning.

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Abstract Background A vaginal fistula is a devastating condition, affecting an estimated 2 million girls and women across Africa and Asia. There are numerous challenges associated with providing fistula repair services in developing countries, including limited availability of operating rooms, equipment, surgeons with specialized skills, and funding from local or international donors to support surgeries and subsequent post-operative care. Finding ways of providing services in a more efficient and cost-effective manner, without compromising surgical outcomes and the overall health of the patient, is paramount. Shortening the duration of urethral catheterization following fistula repair surgery would increase treatment capacity, lower costs of services, and potentially lower risk of healthcare-associated infections among fistula patients. There is a lack of empirical evidence supporting any particular length of time for urethral catheterization following fistula repair surgery. This study will examine whether short-term (7 day) urethral catheterization is not worse by more than a minimal relevant difference to longer-term (14 day) urethral catheterization in terms of incidence of fistula repair breakdown among women with simple fistula presenting at study sites for fistula repair service. Methods/Design This study is a facility-based, multicenter, non-inferiority randomized controlled trial (RCT) comparing the new proposed short-term (7 day) urethral catheterization to longer-term (14 day) urethral catheterization in terms of predicting fistula repair breakdown. The primary outcome is fistula repair breakdown up to three months following fistula repair surgery as assessed by a urinary dye test. Secondary outcomes will include repair breakdown one week following catheter removal, intermittent catheterization due to urinary retention and the occurrence of septic or febrile episodes, prolonged hospitalization for medical reasons, catheter blockage, and self-reported residual incontinence. This trial will be conducted among 512 women with simple fistula presenting at 8 study sites for fistula repair surgery over the course of 24 months at each site. Discussion If no major safety issues are identified, the data from this trial may facilitate adoption of short-term urethral catheterization following repair of simple fistula in sub-Saharan Africa and Asia. Trial registration ClinicalTrials.gov Identifier NCT01428830.

Opening portion of theme section. 8 contributions.

Abstract Background We present a comprehensive technological solution for bacterial diagnostics using tmRNA as a marker molecule. A robust probe design algorithm for microbial detection microarray is implemented. The probes were evaluated for specificity and, combined with NASBA (Nucleic Acid Sequence Based Amplification) amplification, for sensitivity. Results We developed a new web-based program SLICSel for the design of hybridization probes, based on nearest-neighbor thermodynamic modeling. A SLICSel minimum binding energy difference criterion of 4 kcal/mol was sufficient to design of Streptococcus pneumoniae tmRNA specific microarray probes. With lower binding energy difference criteria, additional hybridization specificity tests on the microarray were needed to eliminate non-specific probes. Using SLICSel designed microarray probes and NASBA we were able to detect S. pneumoniae tmRNA from a series of total RNA dilutions equivalent to the RNA content of 0.1-10 CFU. Conclusions The described technological solution and both its separate components SLICSel and NASBA-microarray technology independently are applicative for many different areas of microbial diagnostics.

Abstract Background Despite the implementation of prevention guidelines, early-onset group B streptococci (GBS) disease remains a cause of neonatal morbidity and mortality worldwide. Strategies to identify women who are at risk of transmitting GBS to their infant and the administration of intrapartum antibiotics have greatly reduced the incidence of neonatal GBS disease. However, there is a requirement for a rapid diagnostic test for GBS that can be carried out in a labour ward setting especially for women whose GBS colonisation status is unknown at the time of delivery. We report the design and evaluation of a real-time PCR test (RiboSEQ GBS test) for the identification of GBS in vaginal swabs from pregnant women. Methods The qualitative real-time PCR RiboSEQ GBS test was designed based on the bacterial ssrA gene and incorporates a competitive internal standard control. The analytical sensitivity of the test was established using crude lysate extracted from serial dilutions of overnight GBS culture using the IDI Lysis kit. Specificity studies were performed using DNA prepared from a panel of GBS strains, related streptococci and other species found in the genital tract environment. The RiboSEQ GBS test was evaluated on 159 vaginal swabs from pregnant women and compared with the GeneOhm™ StrepB Assay and culture for the identification of GBS. Results The RiboSEQ GBS test is specific and has an analytical sensitivity of 1-10 cell equivalents. The RiboSEQ GBS test was 96.4% sensitive and 95.8% specific compared to "gold standard" culture for the identification of GBS in vaginal swabs from pregnant women. In this study, the RiboSEQ GBS test performed slightly better than the commercial BD GeneOhm™ StrepB Assay which gave a sensitivity of 94.6% and a specificity of 89.6% compared to culture. Conclusion The RiboSEQ GBS test is a valuable method for the rapid, sensitive and specific detection of GBS in pregnant women. This study also validates the ssrA gene as a suitable and versatile target for nucleic acid-based diagnostic tests for bacterial pathogens.

Abstract Background A real-time multiplex PCR assay was developed for the detection of multiple Salmonella serotypes in chicken samples. Poultry-associated serotypes detected in the assay include Enteritidis, Gallinarum, Typhimurium, Kentucky and Dublin. The traditional cultural method according to EN ISO 6579:2002 for the detection of Salmonella in food was performed in parallel. The real-time PCR based method comprised a pre-enrichment step in Buffered Peptone Water (BPW) overnight, followed by a shortened selective enrichment in Rappaport Vasilliadis Soya Broth (RVS) for 6 hours and subsequent DNA extraction. Results The real-time multiplex PCR assay and traditional cultural method showed 100% inclusivity and 100% exclusivity on all strains tested. The real-time multiplex PCR assay was as sensitive as the traditional cultural method in detecting Salmonella in artificially contaminated chicken samples and correctly identified the serotype. Artificially contaminated chicken samples resulted in a detection limit of between 1 and 10 CFU per 25 g sample for both methods. A total of sixty-three naturally contaminated chicken samples were investigated by both methods and relative accuracy, relative sensitivity and relative specificity of the real-time PCR method were determined to be 89, 94 and 87%, respectively. Thirty cultures blind tested were correctly identified by the real-time multiplex PCR method. Conclusion Real-time PCR methodology can contribute to meet the need for rapid identification and detection methods in food testing laboratories.

Background: Reintegration of women after repair of their female genital fistula remains a challenge. The objective of this study was to document the medical pathway and the reintegration process of women through the "social immersion" program of EngenderHealth in Kissidougou and Labé (Guinea). Methods: This was a qualitative descriptive study with 55 participants, including women seeking fistula care and stakeholders involved in the social immersion for repaired women in Kissidougou and Labé. The study included collecting demographic and clinical data of women, interviews with women before and after surgery, after social immersion, and 3 months post-discharge. Municipal officials, health providers, and members of host families were also interviewed. The study protocol was approved by the Guinea National Ethics Committee for Health Research. Results: The study confirmed that obstetric fistula still occurs among women living in rural and underserved areas. Most women attended at least two to five antenatal care visits, but nine over 10 reported a tragic experience of child loss associated with the occurrence of fistula. Most of them received support from their husband/partner during referral after the obstructed labor and later in the search for treatment. Women and stakeholders reported a good experience of surgery and social immersion in both Kissidougou and Labé. About 3 months after discharge, women who were continent reported being happy with their new life compared to women discharged with repair failure and residual incontinence. Conclusion: The study found the positive impacts of social immersion on the quality of life of women after fistula repair, particularly for those women who had a successful repair. The approach can be included in fistula care programs, either through direct provision or through referral to programs that can provide this service., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Delamou, Douno, Bouédouno, Millimono, Barry, Tripathi and Diallo.)

Background: Female genital fistula is a devastating maternal complication of delivery in developing countries. We sought to analyse the incidence and proportion of fistula recurrence, residual urinary incontinence, and pregnancy after successful fistula closure in Guinea, and describe the delivery-associated maternal and child health outcomes., Methods: We did a longitudinal study in women discharged with a closed fistula from three repair hospitals supported by EngenderHealth in Guinea. We recruited women retrospectively (via medical record review) and prospectively at hospital discharge. We used Kaplan-Meier methods to analyse the cumulative incidence, incidence proportion, and incidence ratio of fistula recurrence, associated outcomes, and pregnancy after successful fistula closure. The primary outcome was recurrence of fistula following discharge from repair hospital in all eligible women who consented to inclusion and could provide follow-up data., Findings: 481 women eligible for analysis were identified retrospectively (from Jan 1, 2012, to Dec 31, 2014; 348 women) or prospectively (Jan 1 to June 20, 2015; 133 women), and followed up until June 30, 2016. Median follow-up was 28·0 months (IQR 14·6-36·6). 73 recurrent fistulas occurred, corresponding to a cumulative incidence of 71 per 1000 person-years (95% CI 56·5-89·3) and an incidence proportion of 18·4% (14·8-22·8). In 447 women who were continent at hospital discharge, we recorded 24 cases of post-repair residual urinary incontinence, equivalent to a cumulative incidence of 23·1 per 1000 person-years (14·0-36·2), and corresponding to 10·3% (5·2-19·6). In 305 women at risk of pregnancy, the cumulative incidence of pregnancy was 106·0 per 1000 person-years, corresponding to 28·4% (22·8-35·0) of these women. Of 50 women who had delivered by the time of follow-up, only nine delivered by elective caesarean section. There were 12 stillbirths, seven delivery-related fistula recurrences, and one maternal death., Interpretation: Recurrence of female genital fistula and adverse pregnancy-related maternal and child health outcomes were frequent in women after fistula repair in Guinea. Interventions are needed to safeguard the health of women after fistula repair., Funding: Belgian Development Cooperation (DGD), Institute of Tropical Medicine of Antwerp (ITM), and Maferinyah Training and Research Center in Rural Health (Guinea)., (Copyright © 2017 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY 4.0 license. Published by Elsevier Ltd.. All rights reserved.)

Background: The prevention and treatment of obstetric fistula still remains a concern and a challenge in low income countries. The objective of this study was to estimate the overall proportions of failure of fistula closure and incontinence among women undergoing repair for obstetric fistula in Guinea and identify its associated factors., Methods: This was a retrospective cohort study using data extracted from medical records of fistula repairs between 1 January 2012 and 30 September 2013. The outcome was the failure of fistula closure and incontinence at hospital discharge evaluated by a dye test. A sub-sample of women with vesicovaginal fistula was used to identify the factors associated with these outcomes., Results: Overall, 109 women out of 754 (14.5 %; 95 % CI:11.9-17.0) unsuccessful repaired fistula at discharge and 132 (17.5 %; 95 % CI:14.8-20.2) were not continent. Failure of fistula closure was associated with vaginal delivery (AOR: 1.9; 95 % CI: 1.0-3.6), partially (AOR: 2.0; 95 % CI: 1.1-5.6) or totally damaged urethra (AOR: 5.9; 95 % CI: 2.9-12.3) and surgical repair at Jean Paul II Hospital (AOR: 2.5; 95 % CI: 1.2-4.9). Women who had a partially damaged urethra (AOR: 2.5; 95 % CI: 1.5-4.4) or a totally damaged urethra (AOR: 6.3; 95 % CI: 3.0-13.0) were more likely to experience post-repair urinary incontinence than women who had their urethra intact., Conclusion: At programmatic level in Guinea, caution should be paid to the repair of women who present with a damaged urethra and those who delivered vaginally as they carry greater risks of experiencing a failure of fistula closure and incontinence.

Objective: To analyze the management of obstetric vesico-vaginal fistula in the three sites of Engender Health in Guinea., Patients and Methods: It was a retrospective study of descriptive type having helped collect 450 cases of vesico-vaginal fistulas in three support sites engender health between January 2008 and December 2011. The variables studied were epidemiological, clinical and therapeutic reasons and treatment outcomes were evaluated after a decline of at least six months., Results: The mean age of onset of the fistula was 25years, ranging from 12 to 55years and 58.8% (n=265) of patients were aged between 18 and 30years. The mean duration of fistula was 11years, ranging from 1 to 38years. Eighty-two percent (n=416) of patients were housewives and 66.4% (n=299) off school. The complex fistula with 66% (n=297) was the most frequent. The treatment consisted of a fistulorraphie after splitting vesico-vaginal in 93.3% (n=420) of cases. Therapeutic results considered after a mean of 8months have resulted in a cure in 79.3% (n=357) of cases, improvement in 4.2% (n=19) of cases and failure in 16 4% (n=74) of cases., Conclusion: Vesico-vaginal fistula is a major cause of maternal morbidity in Guinea. The establishment of a real health policy based on sound medical and social structures contributes to its eradication., Level of Evidence: 5., (Copyright © 2016 Elsevier Masson SAS. All rights reserved.)

Objectives: To analyse the trend of loss to follow-up over time and identify factors associated with women being lost to follow-up after discharge in three fistula repair hospitals in Guinea., Methods: This retrospective cohort study used data extracted from medical records of fistula repairs conducted from 1 January 2007 to 30 September 2013. A woman was considered lost to follow-up if she did not return within 4 months post-discharge. Factors associated with loss to follow-up were identified using a subsample of the data covering the period 2010-2013., Results: Over the study period, the proportion of loss to follow-up was 21.5% (448/2080) and varied across repair hospitals and over time with an increase from 2% in 2009 to 52% in 2013. After adjusting for other variables in a multivariate logistic regression model, women who underwent surgery at Labe hospital and at Kissidougou hospital were more likely to be lost to follow-up than women operated at Jean Paul II hospital (OR: 50.6; 95% CI: 24.9-102.8) and (OR: 11.5; 95% CI: 6.1-22.0), respectively. Women with their fistula closed at hospital discharge (OR: 3.2; 95% CI: 2.1-4.8) and women admitted for repair in years 2011-2013 showed higher loss to follow-up as compared to 2010. Finally, loss to follow-up increased by 2‰ for each additional kilometre of distance a client lived from the repair hospital (OR: 1.002; 95% CI: 1.001-1.003)., Conclusion: Reimbursement of transport was the likely reason for change over time of LTFU. Reducing geographical barriers to care for women with fistula could sustain fistula care positive outcomes., (© 2015 The Authors. Tropical Medicine & International Health Published by John Wiley & Sons Ltd.)

Background: Duration of bladder catheterisation after female genital fistula repair varies widely. We aimed to establish whether 7 day bladder catheterisation was non-inferior to 14 days in terms of incidence of fistula repair breakdown in women with simple fistula., Methods: In this randomised, controlled, open-label, non-inferiority trial, we enrolled patients at eight hospitals in the Democratic Republic of the Congo, Ethiopia, Guinea, Kenya, Niger, Nigeria, Sierra Leone, and Uganda. Consenting patients were eligible if they had a simple fistula that was closed after surgery and remained closed 7 days after surgery, understood study procedures and requirements, and agreed to return for follow-up 3 months after surgery. We excluded women if their fistula was not simple or was radiation-induced, associated with cancer, or due to lymphogranuloma venereum; if they were pregnant; or if they had multiple fistula. A research assistant at each site randomly allocated participants 1:1 (randomly varying block sizes of 4-6; stratified by country) to 7 day or 14 day bladder catheterisation (via a random allocation sequence computer generated centrally by WHO). Outcome assessors were not masked to treatment assignment. The primary outcome was fistula repair breakdown, on the basis of dye test results, any time between 8 days after catheter removal and 3 months after surgery. The non-inferiority margin was 10%, assessed in the per-protocol population. This trial is registered with ClinicalTrials.gov, number NCT01428830., Findings: We randomly allocated 524 participants between March 7, 2012, and May 6, 2013; 261 in the 7 day group and 263 in the 14 day group. In the per-protocol analysis, ten (4%) of 250 patients had repair breakdown in the 7 day group (95% CI 2-8) compared with eight (3%) of 251 (2-6) in the 14 day group (risk difference 0·8% [95% CI -2·8 to 4·5]), meeting the criteria for non-inferiority., Interpretation: 7 day bladder catheterisation after repair of simple fistula is non-inferior to 14 day catheterisation and could be used for management of women after repair of simple fistula with no evidence of a significantly increased risk of repair breakdown, urinary retention, or residual incontinence up to 3 months after surgery., Funding: US Agency for International Development., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Introduction: In 2010, the Ministry of Health (MoH) of Guinea introduced a free emergency obstetric care policy in all the public health facilities of the country. This included antenatal checks, normal delivery and Caesarean section., Objective: This study aims at assessing the changes in coverage of obstetric care according to the Unmet Obstetric Need concept before (2008) and after (2012) the implementation of the free emergency obstetric care policy in a rural health district in Guinea., Methods: We carried out a descriptive cross-sectional study involving the retrospective review of routine programme data during the period April to June 2014., Results: No statistical difference was observed in women's sociodemographic characteristics and indications (absolute maternal indications versus non-absolute maternal indications) before and after the implementation of the policy. Compared to referrals from health centers of patients, direct admissions at hospital significantly increased from 49% to 66% between 2008 and 2012 (p = 0.001). In rural areas, this increase concerned all maternal complications regardless of their severity, while in urban areas it mainly affected very severe complications. Compared to 2008, there were significantly more Major Obstetric Interventions for Maternal Absolute Indications in 2012 (p < 0.001). Maternal deaths decreased between 2008 and 2012 from 1.5% to 1.1% while neonatal death increased from 12% in 2008 to 15% in 2012., Conclusion: The implementation of the free obstetric care policy led to a significant decrease in unmet obstetric need between 2008 and 2012 in the health district of Kissidougou. However, more research is needed to allow comparisons with other health districts in the country and to analyse the trends.

Objectives: Female genital fistula remains a public health concern in developing countries. From January 2007 to September 2013, the Fistula Care project, managed by EngenderHealth in partnership with the Ministry of Health and supported by USAID, integrated fistula repair services in the maternity wards of general hospitals in Guinea. The objective of this article was to present and discuss the clinical outcomes of 7 years of work involving 2116 women repaired in three hospitals across the country., Methods: This was a retrospective cohort study using data abstracted from medical records for fistula repairs conducted from 2007 to 2013. The study data were reviewed during the period April to August 2014., Results: The majority of the 2116 women who underwent surgical repair had vesicovaginal fistula (n = 2045, 97%) and 3% had rectovaginal fistula or a combination of both. Overall 1748 (83%) had a closed fistula and were continent of urine immediately after surgery. At discharge, 1795 women (85%) had a closed fistula and 1680 (79%) were dry, meaning they no longer leaked urine and/or faeces. One hundred and fifteen (5%) remained with residual incontinence despite fistula closure. Follow-up at 3 months was completed by 1663 (79%) women of whom 1405 (84.5%) had their fistula closed and 80% were continent. Twenty-one per cent were lost to follow-up., Conclusion: Routine programmatic repair for obstetric fistula in low resources settings can yield good outcomes. However, more efforts are needed to address loss to follow-up, sustain the results and prevent the occurrence and/or recurrence of fistula., (© 2015 The Authors. Tropical Medicine & International Health Published by John Wiley & Sons Ltd.)

Background: Treatment and care for female genital fistula have become increasingly available over the last decade in countries across Africa and South Asia. Before the International Federation of Gynaecology and Obstetrics (FIGO) and partners published a global fistula training manual in 2011 there was no internationally recognized, standardized training curriculum, including perioperative care. The community of fistula care practitioners and advocates lacks data about the prevalence of various perioperative clinical procedures and practices and their potential programmatic implications are lacking., Methods: Data presented here are from a prospective cohort study conducted between September 2007 and September 2010 at 11 fistula repair facilities supported by Fistula Care in five countries. Clinical procedures and practices used in the routine perioperative management of over 1300 women are described., Results: More than two dozen clinical procedures and practices were tabulated. Some of them were commonly used at all sites (e.g., vaginal route of repair, 95.3% of cases); others were rare (e.g., flaps/grafts, 3.4%) or varied widely depending on site (e.g. for women with urinary fistula, the inter-quartile range for median duration of post-repair bladder catheterization was 14 to 29 days)., Conclusions: These findings show a wide range of clinical procedures and practices with different program implications for safety, efficacy, and cost-effectiveness. The variability indicates the need for further research so as to strengthen the evidence base for fistula treatment in developing countries.

This article presents data from 1354 women from five countries who participated in a prospective cohort study conducted between 2007 and 2010. Women undergoing surgery for fistula repair were interviewed at the time of admission, discharge, and at a 3-month follow-up visit. While women's experiences differed across countries, a similar picture emerges across countries: women married young, most were married at the time of admission, had little education, and for many, the fistula occurred after the first pregnancy. Median age at the time of fistula occurrence was 20.0 years (interquartile range 17.3-26.8). Half of the women attended some antenatal care (ANC); among those who attended ANC, less than 50% recalled being told about signs of pregnancy complications. At follow-up, most women (even those who were not dry) reported improvements in many aspects of social life, however, reported improvements varied by repair outcome. Prevention and treatment programmes need to recognise the supportive role that husbands, partners, and families play as women prepare for safe delivery. Effective treatment and support programmes are needed for women who remain incontinent after surgery.