Use of ATP Readings to Predict a Successful Hygiene Intervention in the Workplace to Reduce the Spread of Viruses on Fomites

The purpose of this study was to validate the use of adenosine triphosphate (ATP) for evaluating hygiene intervention effectiveness in reducing viral dissemination in an office environment. The bacterial virus MS-2 was used to evaluate two scenarios, one where the hand of an individual was contaminated and another where a fomite was contaminated. MS-2 was selected as a model because its shape and size are similar to many human pathogenic viruses. Two separate experiments were conducted, one in which the entrance door push plate was inoculated and the other in which the hand of one selected employee was inoculated. In both scenarios, 54 selected surfaces in the office were tested to assess the dissemination of the virus within the office. Associated surface contamination was also measured employing an ATP meter. More than half of the tested hands and surfaces in the office were contaminated with MS-2 within 4 h. Next, an intervention was conducted, and each scenario was repeated. Half of the participating employees were provided hand sanitizer, facial tissues, and disinfecting wipes, and were instructed in their use. A significant (p < 0.05) reduction was observed in the number of surfaces contaminated with virus. This reduction in viral spread was evident from the results of both viral culture and the surface ATP measurements, although there was no direct correlation between ATP measurements with respect to viral concentration. Although ATP does not measure viruses, these results demonstrate that ATP measurements could be useful for evaluating the effectiveness of hygiene interventions aimed at preventing viral spread in the workplace.     

Norovirus GII.Pe Genotype: Tracking a Foodborne Outbreak on a Cruise Ship Through Molecular Epidemiology,Brazil, 2014

Norovirus (NoV) is recognized as the most common cause of foodborne outbreaks. In 2014, an outbreak of acute gastroenteritis occurred on a cruise ship in Brazil, and NoV became the suspected etiology. Here we present the molecular identification of the NoV strains and the use of sequence analysis to determine modes of virus transmission. Food (cream cheese, tuna salad, grilled fish, orange mousse, and vegetables soup) and clinical samples were analyzed by ELISA, conventional RT-PCR, qRT-PCR, and sequencing. Genogroup GII NoV was identified by ELISA and conventional RT-PCR in fecal samples from 5 of 12 patients tested (41.7%), and in the orange mousse food sample by conventional RT-PCR and qRT-PCR. Two fecal GII NoV samples and the orange mousse GII NoV sample were successfully genotyped as GII.Pe (ORF 1), revealed 98.0–98.8% identities among them, and shared phylogenetically distinct cluster. Establishing the source of a NoV outbreak can be a challenging task. In this report, the molecular analysis of the partial RdRp NoV gene provided a powerful tool for genotyping (GII.Pe) and tracking of outbreak-related samples. In addition, the same fast and simple extraction methods applied to clinical samples could be successfully used for complex food matrices, and have the potential to be introduced in routine laboratories for screening foods for presence of NoV.     

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.     

DNA Heat Treatment for Improving qPCR Analysis of Human Adenovirus in Wastewater

PCR inhibitory substances in complex sample matrices can cause false negatives or under-estimation of target concentration. This study assessed DNA heat treatment for reducing inhibition during qPCR analysis of human adenovirus (HAdV) in wastewater samples. Inhibition was reduced by heat treating DNA, where mean HAdV concentration was increased by 0.71 log₁₀ GC/L (and up to 3.04 log₁₀ GC/L in one case), and replicate variability and false negatives were reduced. DNA heat treatment should be further investigated for improving reliability of HAdV concentration estimates in water, which can support more accurate assessment of health risks associated with viral pathogen exposure.     

Assessment and Evaluation of an Integrated Hybrid Anaerobic–Aerobic Sewage Treatment System for the Removal of Enteric Viruses

The capability of a cost-effective and a small size decentralized pilot wastewater treatment plant (WWTP) to remove enteric viruses such as rotavirus, norovirus genogroup I (GGI), norovirus genogroup II (GGII), Hepatitis E virus (HEV), and adenovirus was studied. This pilot plant is an integrated hybrid anaerobic/aerobic setup which consisted of anaerobic sludge blanket (UASB), biological aerated filter (BAF), and inclined plate settler (IPS). Both the UASB and BAF are packed with a non-woven polyester fabric (NWPF). Results indicated that the overall log₁₀ reductions of enteric viruses’ genome copies through the whole system were 3.1 ± 1, 3.3 ± 0.5, and 2.6 ± 0.9 log₁₀ for rotavirus, norovirus GGI, and adenovirus, respectively. Reduction efficiency for both norovirus GGII and HEV after the different treatment steps could not be calculated because there were no significant numbers of positive samples for both viruses. The overall reduction of rotavirus infectious units through the whole system was 2.2 ± 0.8 log₁₀ reduction which is very close to the overall log₁₀ reduction of adenovirus infectious units through the whole system which was 2.1 ± 0.8 log₁₀ reduction. There was no considerable difference in the removal efficiency for different rotavirus G and P types. Adenovirus 41 was the only type detected in the all positive samples. Although the pilot WWTP investigated is cost effective, has a small footprint, does not need a long distance network pipes, and easy to operate, its efficiency to remove enteric viruses is comparable with the conventional centralized WWTPs.     

典型水库型湖泊中CDOM吸收及荧光光谱变化特征:基于沿岸生态系统分析

为进一步了解水库型湖泊溶解性有机质(DOM)的地球化学特征,本文以三峡库区典型内陆水库型湖泊——长寿湖为研究对象,利用紫外-可见和三维荧光光谱,并结合湖区周边生态系统分析,讨论了长寿湖水体中CDOM的组成、来源和空间分布特征.结果表明,长寿湖不同采样点DOM浓度(DOC和CDOM)存在一定程度的空间分布差异,但各点FDOM分布较为稳定.回水区出现DOM累积,由于受陆源输入影响有限,水体内源活动主导,具有较明显"内源控制"特征,芳香性和分子量相对较低;而周边陆地以人工林兼旅游开发为主的采样点,陆源输入在带入较多腐殖化(高芳香性)组分的同时,人为活动排放也是导致其类蛋白组分丰富的重要原因;入湖区尽管周边果林和居民生活对水体DOM有一定影响,但上游河流输入的影响也不容忽视.另外,各采样点也出现了不受周边生态系统影响的独立的相关性特征,例如芳香性特征常数(SUVA280)和光谱斜率[S(275~295)]显著负相关、CDOM和FDOM极显著正相关、CDOM和S(275~295)负相关等.同时,长寿湖水体中CDOM的生色团主要由具有芳香性结构的大分子组分构成;至少51%的CDOM波动可以通过FDOM变化来进行解释,其中回水区荧光组分对CDOM变动的影响最为明显.在采用传统FI值无法区分DOM来源差异性时,结合采样点沿岸生态系统,综合紫外-可见和荧光光谱特征,有助于对DOM组成及来源进行解析.     

固载化瓜环的制备及其对磺胺间甲氧嘧啶的吸附性能研究

在微波无极灯下将烯氧基瓜环固载到巯基硅胶(MSG)上,制得固载化瓜环(ACB[6]-SG).通过红外光谱表征固载产物,并研究固载化瓜环对磺胺间甲氧嘧啶(SMM)的吸附性能.结果表明,SMM在固载化瓜环上的吸附基本在2 min内即可达到平衡;Henry方程和Freundlich方程均能较好地拟合SMM在固载化瓜环和巯基硅胶上的等温吸附曲线,但是SMM在固载化瓜环上的吸附主要是在瓜环中的分配,而SMM在巯基硅胶上的吸附则主要是多分子层的表面吸附作用;SMM在固载化瓜环上的平衡吸附量远大于在巯基硅胶上的平衡吸附量;SMM在两者上的吸附过程是一个放热过程,温度越低吸附效果越好.