Improvement of the Bag-Mediated Filtration System for Sampling Wastewater and Wastewater-Impacted Waters

Environmental surveillance of poliovirus (PV) plays an important role in the global program for eradication of wild PV. The bag-mediated filtration system (BMFS) was first developed in 2014 and enhances PV surveillance when compared to the two-phase grab method currently recommended by the World Health Organization (WHO). In this study, the BMFS design was improved and tested for its usability in wastewater and wastewater-impacted surface waters in Nairobi, Kenya. Modifications made to the BMFS included the size, color, and shape of the collection bags, the filter housing used, and the device used to elute the samples from the filters. The modified BMFS concentrated 3–10 L down to 10 mL, which resulted in an effective volume assayed (900–3000 mL) that was 6–20 times greater than the effective volume assayed for samples processed by the WHO algorithm (150 mL). The system developed allows for sampling and in-field virus concentration, followed by transportation of the filter for further analysis with simpler logistics than the current methods. This may ultimately reduce the likelihood of false-negative samples by increasing the effective volume assayed compared to samples processed by the WHO algorithm, making the BMFS a valuable sampling system for wastewater and wastewater-impacted surface waters.     

Poliovirus and Other Enteroviruses from Environmental Surveillance in Italy, 2009–2015

Within the initiatives for poliomyelitis eradication by WHO, Italy activated an environmental surveillance (ES) in 2005. ES complements clinical Acute Flaccid Paralysis (AFP) surveillance for possible polio cases, detects poliovirus circulation in environmental sewage, and is used to monitor transmission in communities. In addition to polioviruses, the analyses comprised: (i) the monitoring of the presence of non-polio enteroviruses in sewage samples and (ii) the temporal and geographical distribution of the detected viruses. From 2009 to 2015, 2880 sewage samples were collected from eight cities participating in the surveillance. Overall, 1479 samples resulted positive for enteroviruses. No wild-type polioviruses were found, although four Sabin-like polioviruses were detected. The low degree of mutation found in the genomes of these four isolates suggests that these viruses have had a limited circulation in the population. All non-polio enteroviruses belonged to species B and the most frequent serotype was CV-B5, followed by CV-B4, E-11, E-6, E-7, CV-B3, and CV-B2. Variations in the frequency of different serotypes were also observed in different seasons and/or Italian areas. Environmental surveillance in Italy, as part of the ‘WHO global polio eradication program’, is a powerful tool to augment the polio surveillance and to investigate the silent circulation or the re-emergence of enteroviruses in the population.     

Environmental Surveillance of Polioviruses in Rio de Janeiro,Brazil, in Support to the Activities of Global Polio Eradication Initiative

Wild polioviruses still remain endemic in three countries (Afghanistan, Pakistan, and Nigeria) and re-emergency of wild polio has been reported in previously polio-free countries. Environmental surveillance has been used as a supplementary tool in monitoring the circulation of wild poliovirus (PVs) and/or vaccine-derived PVs even in the absence of acute flaccid paralysis cases. This study aimed to monitor the presence of polioviruses in wastewater samples collected at one wastewater treatment plant located in the municipality of Rio de Janeiro, Brazil. From December 2011 to June 2012 and from September to December 2012, 31 samples were collected and processed. RD and L20B cell cultures were able to isolate PVs and non-polio enteroviruses in 27/31 samples. Polioviruses were isolated in eight samples (type 1 Sabin = 1, type 2 Sabin = 5, and type 3 Sabin = 2). Vaccine-derived polioviruses were not detected nor evidence of recombination with other PVs or non-polio enterovirus serotypes were observed among the isolates. The Sabin-related serotypes 2 and 3 presented nucleotide substitutions in positions associated with the neurovirulent phenotype at the 5′-UTR. Changes in important Amino acid residues at VP1 were also observed in the serotypes 2 and 3. Environmental surveillance has been used successfully in monitoring the circulation of PVs and non-polio enteroviruses and it is of crucial importance in the final stages of the WHO global polio eradication initiative. Our results show the continuous circulation of Sabin-like PVs and non-polio enteroviruses in the analyzed area during the study period.     

Prevalence and Bayesian Phylogenetics of Enteroviruses Derived From Environmental Surveillance Around Polio Vaccine Switch Period in Shandong Province,China

Food and Environmental Virology - We present the results of environmental surveillance for poliovirus (PV) and non-poliovirus (NPEV) around the switch from trivalent to bivalent oral polio-vaccine...     

Use of Preservative Agents and Antibiotics for Increased Poliovirus Survival on Positively Charged Filters

Environmental surveillance of poliovirus (PV) and other non-enveloped viruses can help identify silent circulation and is necessary to certify eradication. The bag-mediated filtration system is an efficient method to filter large volumes of environmental waters at field sites for monitoring the presence of viruses. As filters may require long transit times to off-site laboratories for processing, viral inactivation or overgrowth of bacteria and fungi can interfere with virus detection and quantification (Miki and Jacquet in Aquatic Microb Ecol 51(2):195–208, 2008). To evaluate virus survival over time on ViroCap^? filters, the filters were seeded with PV type 1 (PV1) and/or MS2 and then dosed with preservatives or antibiotics prior to storage and elution. These filters were stored at various temperatures and time periods, and then eluted for PV1 and MS2 recovery quantification. Filters dosed with the preservative combination of 2% sodium benzoate and 0.2% calcium propionate had increased virus survival over time when stored at 25 °C, compared to samples stored at 25 °C with no preservatives. While elution within 24 h of filtration is recommended, if storage or shipping is required then this preservative mixture can help preserve sample integrity. Addition of an antibiotic cocktail containing cephapirin, gentamicin, and Proclin^? 300 increased recovery after storage at 4 and 25 °C, when compared to storage with no antibiotics. The antibiotic cocktail can aid sample preservation if access to appropriate antibiotics storage is available and sample cold chain is unreliable. This study demonstrated that the use of preservatives or antibiotics is a simple, cost-effective method to improve virus detection from ViroCap cartridge filters over time.     

Microbial Source Tracking Analysis Using Viral Indicators in Santa Lucía and Uruguay Rivers,Uruguay

The aim of this study was to determine the origin (human, bovine or porcine) and the concentration of the fecal sources of contamination in waters from Santa Lucía basin and Uruguay River in Uruguay by using host-specific viral markers (adenoviruses and polyomaviruses) as microbial source tracking (MST). Between June 2015 and May 2016, monthly collections of surface water samples were performed in six sites in Santa Lucía basin and four sites in Uruguay River (n = 120 samples). Viral concentration was carried out using an absorption-elution method. Detection and quantification of human and porcine adenovirus (HAdV and PAdV, respectively) and human and bovine polyomavirus (HPyV and BoPyV, respectively) were performed by quantitative PCR (qPCR). To evaluate the infectivity of circulating HAdV, an integrated cell culture-qPCR (ICC-qPCR) was used. A logistic regression analysis was carried out to estimate the influence of environmental variables on the virus presence in surface waters. Overall, HAdV was the prevalent (18%; 21/120) followed by BoPyV (11%; 13/120) and HPyV (3%; 3/120), whereas PAdV was not detected in this study. The mean concentration ranged from 1.5 × 10⁴ genomic copies/L (gc/L) for HAdV to 1.8 × 10² gc/L for HPyV. Infective HAdVs were observed in two out of ten analyzed samples. A significant effect of environmental temperature (p = 0.001) and river (p = 0.012) on the presence of human viruses was found. These results suggest that fecal contamination could affect the water quality of these rivers, showing deficiencies in the procedure of sewage discharge from regional cities, livestock and dairy farms.

     

Insights from a Systematic Search for Information on Designs,Costs, and Effectiveness of Poliovirus Environmental Surveillance Systems

Poliovirus surveillance plays a critical role in achieving and certifying eradication and will play a key role in the polio endgame. Environmental surveillance can provide an opportunity to detect circulating polioviruses prior to the observation of any acute flaccid paralysis cases. We completed a systematic review of peer-reviewed publications on environmental surveillance for polio including the search terms “environmental surveillance” or “sewage,” and “polio,” “poliovirus,” or “poliomyelitis,” and compared characteristics of the resulting studies. The review included 146 studies representing 101 environmental surveillance activities from 48 countries published between 1975 and 2016. Studies reported taking samples from sewage treatment facilities, surface waters, and various other environmental sources, although they generally did not present sufficient details to thoroughly evaluate the sewage systems and catchment areas. When reported, catchment areas varied from 50 to over 7.3 million people (median of 500,000 for the 25% of activities that reported catchment areas, notably with 60% of the studies not reporting this information and 16% reporting insufficient information to estimate the catchment area population size). While numerous studies reported the ability of environmental surveillance to detect polioviruses in the absence of clinical cases, the review revealed very limited information about the costs and limited information to support quantitative population effectiveness of conducting environmental surveillance. This review motivates future studies to better characterize poliovirus environmental surveillance systems and the potential value of information that they may provide in the polio endgame.     

Detection of Norovirus Recombinant GII.2[P16] Strains in Oysters in Thailand

Human norovirus causes sporadic and epidemic acute gastroenteritis worldwide, and the predominant strains are genotype GII.4 variants. Recently, a novel GII.17[P17] and a recombinant GII.2[P16] strain have been reported as the causes of gastroenteritis outbreaks. Outbreaks of norovirus are frequently associated with foodborne illness. In this study, each of 75 oyster samples processed by a proteinase K extraction method and an adsorption-elution method were examined for noroviruses using RT-nested PCR with capsid primers. Thirteen (17.3%) samples processed by either method tested positive for norovirus genogroup II (GII). PCR amplicons were characterized by DNA sequencing and phylogenetic analysis as GII.2 (n?=?6), GII.4 (n?=?1), GII.17 (n?=?3), and GII.unclassified (n?=?3). Norovirus-positive samples were further amplified by semi-nested RT-PCR targeting the polymerase-capsid genes. One nucleotide sequence revealed GII.17[P17] Kawasaki strain. Five nucleotide sequences were identified as belonging to the recombinant GII.2[P16] strains by recombination analysis. The collected oyster samples were quantified for norovirus GII genome copy number by RT-quantitative PCR. Using the proteinase K method, GII was found in 13/75 (17.3%) of samples with a range of 8.83–1.85?×?10⁴ genome copies/g of oyster. One sample (1/75, 1.3%) processed by the adsorption-elution method was positive for GII at 5.00?×?10¹ genome copies/g. These findings indicate the circulation of a new variant GII.17 Kawasaki strain and the recombinant GII.2[P16] in oyster samples corresponding to the circulating strains reported at a global scale during the same period of time. The detection of the recombinant strains in oysters emphasizes the need for continuing systematic surveillance for control and prevention of norovirus gastroenteritis.

     

Factors associated with multiple-pass procedures during chorionic villus sampling: A video analysis

Multiple placental passes during chorionic villus sampling (CVS) increase the risk of fetal loss; however, specific factors that predispose to repeat aspiration have not been delineated. To identify anatomic and technical variables associated with multiple-pass procedures, a detailed review of 205 videotaped CVS procedures (single pass = 163; multiple pass = 42) was performed, blinded to pregnancy outcome. The route of sampling did not influence the need for multiple aspiration attempts (transabdominal—30/ 135; transcervical—12/70), nor was placental location alone discriminatory. However, the combination of a posterior placenta and uterine retroversion was observed more frequently in the multiple-pass cohort (8/42 vs. 9/163; p<0.05). In transabdominal cases, suboptimal needle placement (e.g., perpendicular to the placental long axis) was more common in the initial aspiration of a multiple-pass procedure (21/30 vs. 38/105;p<0.01), while limited penetration of the catheter tip (e.g., just inside the placental edge) characterized a majority of multiple-pass cases in the transcervical subset (7/12 vs. 3/58; p<0.0001). A case-control cohort was constructed to evaluate the impact of these technical variables on sampling efficacy, independent of the influence of uterine position and placental site. In that analysis, suboptimal location and/or orientation of the sampling device remained characteristic of multiple-pass cases. We conclude that further reduction in the frequency of multiple-pass procedures might be achieved by consistent placement of the device tip in the central placental mass. Unlike amniocentesis, where any point of amnion entry will suffice, this technical nuance should be emphasized with CVS to maximize the single-pass success rate.     

A cross-country comparison of pregnant women's decision-making and perspectives when opting for non-invasive prenatal testing in the Netherlands and Belgium

Background

The Netherlands and Belgium have been among the first countries to offer non-invasive prenatal testing (NIPT) as a first-tier screening test. Despite similarities, differences exist in counseling modalities and test uptake. This study explored decision-making and perspectives of pregnant women who opted for NIPT in both countries.

Methods

A questionnaire study was performed among pregnant women in the Netherlands (NL) (n = 587) and Belgium (BE) (n = 444) opting for NIPT, including measures on informed choice, personal and societal perspectives on trisomy 21, 18 and 13 and pregnancy termination.

Results

Differences between Dutch and Belgian women were shown in the level of informed choice (NL: 83% vs. BE: 59%, p < 0.001), intention to terminate the pregnancy in case of confirmed trisomy 21 (NL: 51% vs. BE: 62%, p = 0.003) and trisomy 13/18 (NL: 80% vs. BE: 73%, p = 0.020). More Belgian women considered trisomy 21 a severe condition (NL: 64% vs. BE: 81%, p < 0.001). Belgian women more frequently indicated that they believed parents are judged for having a child with trisomy 21 (BE: 42% vs. NL: 16%, p < 0.001) and were less positive about quality of care and support for children with trisomy 21 (BE: 23% vs. NL: 62%, p < 0.001).

Conclusion

Differences in women's decision-making regarding NIPT and the conditions screened for may be influenced by counseling aspects and country-specific societal and cultural contexts.     

Determination of polychlorinated biphenyl congeners in environmental samples

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好氧-厌氧两相堆肥过程中抗生素耐药基因的变化特征及影响因素研究

针对堆肥处理过程中抗生素耐药基因（antibiotic resistance genes,ARGs）去除效率不高、诱导潜在致病菌传播扩散等环境安全问题,采用好氧-厌氧两相堆肥处理猪粪、秸秆,探讨ARGs丰度及微生物群落的变化特征,并明确影响ARGs相对丰度变化的关键因子.结果表明:与单一好氧相比,好氧-厌氧两相堆肥能更有效地降低部分ARGs及潜在致病菌的相对丰度,其中ermB、tetK的相对丰度分别降低了39.68%、72.19%,假单胞菌属（Pseudomonas）、梭菌属（Clostridium_sensu_stricto_1）的相对丰度分别降低了96.09%、90.07%.主成分分析（Principal Component Analysis,PCA）及冗余分析（Redundancy analysis,RDA）得出,微生物群落和环境因子均显著影响ARGs相对丰度的变化（P < 0.05）,贡献率分别为82.0%、71.2%.其中,梭菌属（Clostridium_sense_stricto_1）、黄杆菌属（Flavobacterium）、尿素芽孢杆菌属（Ureibacillus）和海洋杆菌属（Oceanobacillus）是影响ARGs相对丰度变化的主要微生物菌属;温度、电导率、含水率和pH是影响ARGs相对丰度变化的关键环境因子,贡献率分别为22.0%、16.7%、16.2%和15.0%.研究显示:与好氧堆肥相比,好氧-厌氧两相堆肥能更有效地去除ARGs,并抑制潜在致病菌的繁殖扩散,从而减少环境风险;然而部分ARGs和潜在致病菌仍存在增殖现象,因此亟需进一步优化好氧-厌氧两相堆肥工艺.      

水稻土对磷的吸持能力及环境风险研究

以湖南Cd污染区水稻土为供试土壤,研究了土壤活性Al(Al_(ox))、活性Fe(Fe_(ox))、磷吸持指数(PSI)、吸持饱和度(DPSS)和释放风险指数(ERI)及其相关性关系.结果表明:供试水稻土中Al_(ox)和Fe_(ox)含量变异性较大,分别为55.67~1495.90 mg·kg~(-1)和1.50~13.63 g·kg~(-1),但有效磷含量变化幅度不大.水稻土吸持磷能力较弱,全部样品中PSI均低于30,但水稻土磷的吸持饱和度较低,85%的土壤样品DPSS低于磷淋失临界值(15%),且磷释放失风险(ERI)较小.水稻土中Al_(ox)含量和Fe_(ox)含量与PSI存在显著正相关关系(p0.05),而与DPSS之间表现为显著负相关关系(p0.01),DPSS和PSI呈现为显著负相关关系(p0.01).     

渗透汽化膜分离技术在环保中的应用

渗透汽化膜分离技术 (PV)和传统的分离技术相比 ,它具有技术可靠、工艺简单、操作方便及费用经济等优点。主要应用于有机溶剂脱水、有机混合物的分离等 ,并部分实现了工业化。随着高分子科学以及膜合成技术的发展 ,PV逐渐应用到一些新的领域。主要介绍了渗透汽化膜分离的基本原理 ,以及它在环保领域的新应用。     

Derivated fetal haemoglobin as a marker for red cell age in the human fetus reflecting stimulated or impaired red blood cell production

We have determined whether derivated fetal haemoglobin (dHbF, consisting of glycated and acetylated HbF) can be used as a cell age marker for fetal red blood cells (RBCs). Cord blood was obtained between 19 and 39 weeks of gestation from 28 alloimmunised anaemic fetuses (23 RhD+ and 5 Kell) and from 20 non-anaemic fetuses and newborns (controls). Density gradient centrifugation was applied to 36 samples (20 RhD+, 15 controls and 1 Kell) to obtain fractions of increasing cell age. Blood samples were used for measurements of mean cellular volume (MCV), mean cell haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), pyruvate kinase activity (PK) and derivated fetal haemoglobin (dHbF) by cation-exchange HPLC. Reticulocytes were counted only in the whole blood samples. In all density gradient separated RBC fractions, the values for MCV, MCH and PK activity decreased and those of MCHC and dHbF increased with increasing density (equivalent to increasing cell age). The mean density was lower for RBCs of the anaemic RHD group (1.072±0.007 g/ml) than for the non-anaemic controls (1.077±0.005 g/ml) (p<0.05) The RBC density of the Kell sensitised fetus did not differ from those of the controls. In the control group, the values of the cell age markers in whole blood changed significantly with the gestational age, showing an increase of mean age of the erythrocyte population. The best linear relationship was found for dHbF (y=6.28+0.17*weeks; r=0.84; p<0.001). In the anaemic RhD+ fetuses, the RBC age markers did not change with gestational age; the dHbF percentages were lower, and the MCV, MCH, PK values and the reticulocyte counts were higher than in the controls (0.05<p<0.001). The dHbF values of the Kell sensitised fetuses were above (p<0.01) and the reticulocyte counts were below normal (p<0.05) for gestational age. For the anaemic fetuses, a significant number of the dHbF values (86%) and of the reticulocyte counts (78%) differed from the values of the controls (p<0.01). The dHbF percentages in RhD+ fetuses showed the best correlation with the Hb deficit, which is a measure for anaemia (r=−0.81, p<0.0001). We conclude that the percentage derivated HbF may indicate whether the RBC production is normal for gestational age. It may in that sense reflect stimulated or impaired erythropoiesis in alloimmunised haemolytic anaemia. Copyright © 2001 John Wiley & Sons, Ltd.     

纳米银在环境水样和实验介质中的理化性质和毒性效应

纳米银在自然环境中的理化性质可能严重影响其环境效应,本文的目的是为了比较纳米银在自然水样和实验介质条件下的理化性质和毒性效应.实验表征了粒径4 nm表面包裹物为油胺的纳米银,在不同介质条件下的粒径和团聚性,并进一步研究了不同环境介质条件下纳米银暴露对枯草芽孢杆菌的毒性效应.透射电镜照片显示油胺包裹的4 nm纳米银在环境介质中发生明显团聚.油胺包裹的4 nm纳米银暴露会抑制培养基中细菌的生长速率,降低生理盐水溶液中细菌的存活率.纳米银在实验条件下对细菌的毒性呈一定剂量效应关系,但是在环境水样中剂量效应关系不明显.低浓度纳米银在环境水样中的毒性比其在实验介质中弱.纳米银的透析袋暴露实验中,细菌的存活率有显著提升,揭示纳米银与细菌的直接相互作用对毒性效应有重要作用.与实验条件的培养基和生理盐水介质相比,环境水样中的纳米银更趋向以团聚形式存在,与细菌的相互作用几率减小,因此纳米银在环境介质中对细菌的毒性会降低.     

Somatic Coliphage Profiles of Produce and Environmental Samples from Farms in Northern México

Somatic coliphages were quantified in 459 produce and environmental samples from 11 farms in Northern Mexico to compare amounts of somatic coliphages among different types of fresh produce and environmental samples across the production steps on farms. Rinsates from cantaloupe melons, jalapeño peppers, tomatoes, and the hands of workers, soil, and water were collected during 2011–2012 at four successive steps on each farm, from the field before harvest through the packing facility, and assayed by FastPhage MPN Quanti-tray method. Cantaloupe farm samples contained more coliphages than jalapeño or tomato (p range <0.01–0.03). Across production steps, jalapeños had higher coliphage percentages before harvest than during packing (p = 0.03), while tomatoes had higher coliphage concentrations at packing than all preceding production steps (p range <0.01–0.02). These findings support the use of targeted produce-specific interventions at multiple points in the process of growing and packing produce to reduce the risk of enteric virus contamination and improve food safety during fruit and vegetable production.     

Highly sensitive flow-biosensor for toxic phenolic compounds using tyrosinase and acridine orange-adsorbed carbon felt

Tyrosinase (TYR, EC 1.14.18.1) was physically adsorbed onto a carbon felt (CF) together with acridine orange (AO). Coadsorption of AO was essential to prevent the denaturation of the TYR at the CF surface. The resulting TYR and AO-coadsorbed CF (TYR/AO-CF) was successfully utilized as a detection unit of novel and highly sensitive amperometric flow-biosensor for toxic chlorophenol compounds. Standard solutions of phenolic compounds (200 μL) were injected, and the cathodic peak currents due to the reduction current of o-quinones produced by the TYR-catalyzed oxidation (phenolase activity) were detected at the applied potential of ?50 mV vs. Ag/AgCl. In this reaction, the electrochemically generated catechol compounds from o-quinones are re-oxidized repeatedly by catecholase activity of the TYR, leading to a sufficient amplified signal. The TYR/AO-CF exhibited much higher selectivity toward p-chlorophenol as compared with other chlorophenol compounds. When 0.1 mol/L phosphate buffer (pH 7.0) was used as a carrier at flow rate of 3.0 mL/min, cathodic peaks for p-chlorophenol was linear in the concentration range between 0.1 and 10 [xmol/L (sensitivity: 1.41(mA-L)/mmol) with sampling rate (30 samples/h), and the detection limit ofp-chlorophenol was found to be 2.13 ? 10⁸ mol/L (S/N = 3. The ratio of signal and noise is 3). The TYR/AO-CF kept more than 80% of original activity after the storage in 0.1 mol/L phosphate buffer (pH 7.0) containing 0.2 mmol/L AO at 4°C.     

Climate change implications for the nest site selection process and subsequent hatching success of a green turtle population

Sandy beach habitat where sea turtles nest will be affected by multiple climate change impacts. Before these impacts occur, knowledge of how nest site selection and hatching success vary with beach microhabitats is needed to inform managers on how to protect suitable habitats and prepare for scientifically valid mitigation measures at beaches around the world. At a highly successful green turtle (Chelonia mydas) rookery at Akumal, Quintana Roo, Mexico, we measured microhabitat characteristics along the beach crawl (rejected sites) and related nest site conditions (selected sites) to subsequent hatching success rates for 64 nesting events. To our knowledge, this is the first study to report environmental data along the nesting crawl for a green turtle population and the first to use natural breaks in the data to describe their preferred habitat ranges. Our results indicate that turtles were likely using a combination of cues to find nest sites, mainly higher elevations and lower sand surface temperatures (Kruskal-Wallis test, H?=?19.84, p?<?0.001; H?=?10.78, p?<?0.001). Hatching success was significantly and negatively correlated to sand temperature at cloaca depth (Spearman’s ρ?=??0.27, p?=?0.04). Indeed, the preferred range for cloaca sand temperatures at the nest site (26.3–27.5 °C) had significantly higher hatching success rates compared to the highest temperature range (Tukey HSD?=?0.47, p?=?0.05). Sand temperatures at various depths were intercorrelated, and surface and cloaca depth sand temperatures were correlated to air temperature (ρ?=?0.70, p?=?0.00; ρ?=?0.26, p?=?0.04). Therefore, rising air temperatures could alter sand temperature cues for suitable nest sites, preferred nest site ranges, and produce uneven sex ratios or lethal incubating temperatures. Elevation cues and preferred ranges (1.4–2.5 m) may also be affected by sea level rise, risking inundation of nests.