Comparison of four DNA extraction and three preservation protocols for the molecular detection and quantification of soil-transmitted helminths in stool

Background : A DNA extraction and preservation protocol that yields sufficient and qualitative DNA is pivotal for the success of any nucleic acid amplification test (NAAT), but it still poses a challenge for soil-transmitted helminths (STHs), including Ascaris lumbricoides, Trichuris trichiura and the two hookworms (Necator americanus and Ancylostoma duodenale). In the present study, we assessed the impact of different DNA extraction and preservativation protocols on STH-specific DNA amplification from stool. Methodology and principal findings : In a first experiment, DNA was extracted from 37 stool samples with variable egg counts for T. trichiura and N. americanus applying two commercial kits, both with and without a prior bead beating step. The DNA concentration of T. trichiura and N. americanus was estimated by means of qPCR. The results showed clear differences in DNA concentration across both DNA extraction kits, which varied across both STHs. They also indicated that adding a bead beating step substantially improved DNA recovery, particularly when the FECs were high. In a second experiment, 20 stool samples with variable egg counts for A. lumbricoides, T. trichiura and N. americanus were preserved in either 96% ethanol, 5% potassium dichromate or RNA later and were stored at 4 degrees C for 65, 245 and 425 days. DNA was extracted using the DNeasy Blood & Tissue kit with a bead beating step. Stool samples preserved in ethanol proved to yield higher DNA concentrations as FEC increased, although stool samples appeared to be stable over time in all preservatives. Conclusions : The choice of DNA extraction kit significantly affects the outcome of NAATs. Given the clear benefit of bead beating and our validation of ethanol for (long-term) preservation, we recommend that these aspects of the protocol should be adopted by any stool sampling and DNA extraction protocol for downstream NAAT-based detection and quantification of STHs

Effect of intermittent aeration cycle on nutrient removal and microbial community in a fluidized bed reactor-membrane bioreactor combo system

Effect of intermittent aeration cycle (IAC=15/45-60/60min) on nutrient removal and microbial community structure was investigated using a novel fluidized bed reactor-membrane bioreactor (FBR-MBR) combo system. FBR alone was found more efficient for removing PO4-P (>85%) than NH4-N (98%). Efficient nitrification, stable mixed liquor suspended solid and reduced transmembrane pressure was also achieved. Quantitative real-time polymerase chain reaction results of total bacteria 16S rRNA gene copies per mL of mixed-liquor varied from (2.48±0.42)×109 initial to (2.74±0.10)×108, (6.27±0.16)×109 and (9.17±1.78)×109 for 15/45, 45/15 and 60/60min of IACs, respectively. The results of clone library analysis revealed that Proteobacteria (59%), Firmicutes (12%) and Bacteroidetes (11%) were the dominant bacterial group in all samples. Overall, the combo system performs optimum nutrient removal and host stable microbial communities at 45/15min of IAC. © 2014 Elsevier Ltd

Lung function reduction and chronic respiratory symptoms among workers in the cement industry: a follow up study

<p>Abstract</p> <p>Background</p> <p>There are only a few follow-up studies of respiratory function among cement workers. The main aims of this study were to measure total dust exposure, to examine chronic respiratory symptoms and changes in lung function among cement factory workers and controls that were followed for one year.</p> <p>Methods</p> <p>The study was conducted in two cement factories in Ethiopia. Totally, 262 personal measurements of total dust among 105 randomly selected workers were performed. Samples of total dust were collected on 37-mm cellulose acetate filters placed in closed faced Millipore-cassettes. Totally 127 workers; 56 cleaners, 44 cement production workers and 27 controls were randomly selected from two factories and examined for lung function and interviewed for chronic respiratory symptoms in 2009. Of these, 91 workers; 38 cement cleaners (mean age 32 years), 33 cement production workers (36 years) and 20 controls (38 years) were examined with the same measurements in 2010.</p> <p>Results</p> <p>Total geometric mean dust exposure among cleaners was 432 mg/m³. The fraction of samples exceeding the Threshold Limit Value (TLV) of 10 mg/m³for the cleaners varied from 84-97% in the four departments. The levels were considerably lower among the production workers (GM = 8.2 mg/m³), but still 48% exceeded 10 mg/m³.</p> <p>The prevalence of all the chronic respiratory symptoms among both cleaners and production workers was significantly higher than among the controls.</p> <p>Forced Expiratory Volume in one second (FEV₁) and FEV₁/Forced Vital Capacity (FEV₁/FVC) were significantly reduced from 2009 to 2010 among the cleaners (p < 0.002 and p < 0.004, respectively) and production workers (p < 0.05 and p < 0.02, respectively), but not among the controls.</p> <p>Conclusions</p> <p>The high prevalence of chronic respiratory symptoms and reduction in lung function is probably associated with high cement dust exposure. Preventive measures are needed to reduce the dust exposure.</p

Patent infections with soil-transmitted helminths and Schistosoma mansoni are not associated with increased prevalence of antibodies to the Onchocerca volvulus peptide epitopes OvMP-1 and OvMP-23

Background: Ov16 serology is considered a reference method for Onchocerca volvulus epidemiological mapping. Given the suboptimal sensitivity of this test and the fact that seroconversion takes more than a year after infection, additional serological tests might be needed to guide onchocerciasis elimination programmes. Recently, two linear epitopes encoded in OvMP-1 and OvMP-23 peptides were introduced as serological markers, but the observed antibody cross-reactivity in samples originating from Onchocerca volvulus non-endemic areas required further investigation. Methods: We evaluated both peptide markers in an O. volvulus hypo-endemic setting in Jimma Town, Ethiopia using peptide ELISA. For all individuals (n = 303), the infection status with soil-transmitted helminths and Schistosoma mansoni was known. Results: We found that 11 (3.6%) individuals were positive for anti-Ov16 IgG4 antibodies, while 34 (11.2%) and 15 (5.0%) individuals were positive for OvMP-1 and OvMP-23, respectively. Out of the 34 OvMP-1 positive samples, 33 were negative on the Ov16 IgG4 ELISA. Similarly, out of the 15 OvMP-23 positive samples, 14 scored negative on this reference method. No difference in seroprevalence for all three markers could be observed between uninfected individuals and individuals infected with different soil-transmitted helminths or S. mansoni. Moreover, the intensity of the response to OvMP-1, OvMP-23 or Ov16 was not significantly stronger in individuals carrying patent STH or S. mansoni infections, nor was there any correlation between the intensities of the responses to the three different antigens. Conclusions: This study demonstrates that a patent infection with either soil-transmitted helminths or S. mansoni does not lead to increased antibody recognition of both OvMP-1 and OvMP23

'We identify, discuss, act and promise to prevent similar deaths': a qualitative study of Ethiopia's Maternal Death Surveillance and Response system.

INTRODUCTION: Ethiopia introduced national Maternal Death Surveillance and Response (MDSR) in 2013 and is among the first sub-Saharan African countries to capture data on facility-based and community-based maternal deaths. We interviewed frontline MDSR implementers about their experiences of the first 2 years of MDSR, including perceptions of its introduction and outcomes for health services. METHODS: We conducted a qualitative case study in 4 zones in the largest regions, interviewing 69 key informants from regional, zonal, district and facility levels. RESULTS: A defining feature of Ethiopia's MDSR system is its integration within existing disease surveillance, with both benefits and challenges. Facilitators of the system's introduction were strong political support, alignment with broader health strategies and strong links across health system departments. Barriers included confusion around new responsibilities, high staff turnover and fear of legal repercussions. Stakeholders believed MDSR increased confidence in using local data to improve maternal health services and enhanced communication across the health system. CONCLUSIONS: MDSR systems take time to establish, encountering challenges in early implementation. Ensuring MDSR has a clear purpose, explicitly defined roles and responsibilities, and adequate supervisory support from the start will ensure it becomes embedded within the health system as routine practice rather than perceived as a stand-alone system. Countries planning to adopt or extend MDSR can learn from Ethiopia's experience, particularly the decision to make maternal mortality a weekly reportable condition within Public Health Emergency Management

Electrocatalytic reduction of carbon dioxide on gold–copper bimetallic nanoparticles: Effects of surface composition on selectivity

In this work, bimetallic nanoparticles (NPs) of Au and Cu are synthesized for the electrochemical reduction reaction of CO2 (CO2RR). It is known that the binding strength between the reduction intermediates and the electrocatalyst affects the selectivity of products, but how to correlate the performance with the surface composition, structure, and properties of a bimetallic electrocatalyst, instead of stoichiometric or bulk composition remains less discussed. AuCu and AuCu3 NPs with the size around similar to 8 nm were prepared. By excluding the size effect, the work studies the effects of surface composition and heteroatomic interaction on the selectivity and faradaic efficiency of the reduction products. Based on X-ray absorption spectroscopy (XAS), one can quantify the alloying extent and surface compositions of electrocatalysts, which are supposed to have the pivotal effects on the reaction pathways and the corresponding reduction products. It is found that high activity and notably improved CO selectivity of Au-Cu bimetallic NPs can be attributed to the heterometallic coordination and their electronic interactions. The reduction products were analyzed by gas chromatography and nuclear magnetic resonance (NMR) spectroscopy. Nearly 60 mA cm(-2) of current density was recorded at -0.91 V vs. RHE, and selectivity of 78 +/- 4.3% CO was obtained using AuCu. (C) 2020 Published by Elsevier Ltd