Antibody- and aptamer-free SERS substrate for ultrasensitive and anti-interference detection of SARS-CoV-2 spike protein in untreated saliva.

Sensitive and anti-interference detection of targeted signal(s) in body fluids is one of the paramount tasks in biosensing. Overcoming the complication and high cost of antibody/aptamer-modification, surface-enhanced Raman spectroscopy (SERS) based on antibody/aptamer-free (AAF) substrates has shown great promise, yet with rather limited detection sensitivity. Herein, we report ultrasensitive and anti-interference detection of SARS-CoV-2 spike protein in untreated saliva by an AAF SERS substrate, applying the evanescent field induced by the high-order waveguide modes of well-defined nanorods for SERS for the first time. A detection limit of 3.6 × 10-17 M and 1.6 × 10-16 M are obtained in phosphate buffered saline and untreated saliva, respectively; the detection limits are three orders of magnitude improved than the best records from AAF substrates. This work unlocks an exciting path to design AAF SERS substrates for ultrasensitive biosensing, not limited to detection of viral antigens.

Alcohol use during COVID-19 in adults with severe untreated AUD.

Evidence demonstrating increased alcohol use during COVID-19 comes from low- to moderate-alcohol use samples and has yet to use adults with severe but untreated AUD. Using a community sample of adults with severe AUD, this exploratory, cross-sectional study examined associations of COVID-19 alcohol use. Participants were recruited for a phase-II RCT. Only baseline measures, completed prior to randomization, were analyzed in the present study. Key variables were alcohol consumption, COVID-19-related worries and experiences, and qualitative responses of 1) alcohol use and 2) positive changes during COVID-19. 176 pariticpants recruited since COVID-19 were on average 41.4 years old, 49.1% female, and 79% White. Participants drank alcohol nearly 23 of the past 30 days, consumed 7 standard drinks per drinking day, and nearly 90% reported increased alcohol use. More heavy episodic drinking was reported in the first six-months of COVID-19 and more COVID-related concerns in the most recent six-months. Participants reported drinking increased due to "more time on their hands", but the pandemic also "strengthened relationships". Results affirm an increase in alcohol use during COVID-19 in adults with severe, untreated AUD. Findings underscore the need to understand how alcohol use and pandemic-related circumstances may influence one another for adults with severe AUD.

Evaluating the transmission risk of SARS-CoV-2 from sewage pollution.

The presence of SARS-CoV-2 in untreated sewage has been confirmed in many countries but its incidence and infection risk in contaminated waters is poorly understood. The River Thames in the UK receives untreated sewage from 57 Combined Sewer Overflows (CSOs), with many discharging dozens of times per year. This study investigated if such discharges provide a pathway for environmental transmission of SARS-CoV-2. Samples of wastewater, surface water, and sediment collected close to six CSOs on the River Thames were assayed over eight months for SARS-CoV-2 RNA and infectious virus. Bivalves were also sampled as an indicator species of viral bioaccumulation. Sediment and water samples from the Danube and Sava rivers in Serbia, where raw sewage is also discharged in high volumes, were assayed as a positive control. No evidence of SARS-CoV-2 RNA or infectious virus was found in UK samples, in contrast to RNA positive samples from Serbia. Furthermore, this study shows that infectious SARS-CoV-2 inoculum is stable in Thames water and sediment for <3 days, while SARS-CoV-2 RNA is detectable for at least seven days. This indicates that dilution of wastewater likely limits environmental transmission, and that detection of viral RNA alone is not an indication of pathogen spillover.

What should General Adult Psychiatrists know about Reproductive Counselling of Women with Severe Mental Illness?

A notable part of psychiatry is prevention. Our job, as psychiatrists, is not only to treat mental disorders, but also to prevent them. Treating mothers in postnatal period reducing the negative consequence of depression for child development, for example. General adult psychiatrists face the challenge of having patients, who receive psychiatric health care, while also planning to have a child. Many professionals may find themselves in a situation, when they feel clueless on what to advise regarding pharmacotherapy and realistic expectations on having a child. The presentation will focus on some crucial topics. What should counselling include when planning pregnancy, highlighting differences among first and second or further children. Potentials risks and harms on the fetus / newborn baby will be introduced with emphasis on pharmacological/ chemical agents, infectious effects and social, relational and family stressors. What the guidelines are for Covid-19 vaccination and pregnancy. Relative and absolute contraindications of planned pregnancy will be discussed. The crucial question of artificial/ therapeutic abortion;are there any psychiatric conditions, when a psychiatrist can/should suggest it? The advantage and knowledge of perinatal mental health guideline papers, bio-ethical aspects will be discussed, along with the consequences of untreated mental illness. Advantage of breastfeeding and an up-to-date view on what should be psychiatrists' aim will be introduced. Why is it inevitable to cooperate with GPs, obstetric- and gynecology colleagues and further medical professionals? The essential aspect and advantages of involving fathers and extended family members in this approach also will be explained.

How to think politically about epidemics: a retrospective and a rejoinder

Cholera – a bacterial infection of the intestine that is easy to prevent but deadly when left untreated – emerges and thrives where modern water and sanitation services have broken down, where people live in squalid and overcrowded conditions, where immune systems are weakened by malnutrition or concomitant infection. For Reddy, this argument underplays the limits of humanitarian responses to achieve more development-oriented transformation. [...]she points out that I don’t give quite enough analysis to the historical role that international organizations have played in Zimbabwe. For Muinde, my book shows how the cholera epidemic shaped new experiences of citizenship among Harare’s urban poor: it tapped into historical expectations of public service delivery, it provided an avenue for township residents to air outrage at the structural inequality in the city and at their political abjection, and it channelled fears about an increasingly fragile future.

Wastewater-based epidemiology in countries with poor wastewater treatment - Epidemiological indicator function of SARS-CoV-2 RNA in surface waters.

Wastewater-based epidemiology (WBE) surveillance of COVID-19 and other future outbreaks is a challenge for developing countries as most households are not connected to a sewerage system. In December 2019, SARS-CoV-2 RNA was detected in the Danube River at a site severely affected by wastewaters from Belgrade. Rivers are much more complex systems than wastewater systems, and efforts are needed to address all the factors influencing the adoption of WBE as an alternative to targeting raw wastewater. Our objective was to provide a more detailed insight into the potential of SARS-CoV-2 surveillance in Serbian surface waters for epidemiological purposes. Water samples were collected at 12 sites along the Sava and Danube rivers in Belgrade during the fourth COVID-19 wave in Serbia that started in late February 2021. RNA was concentrated using Amicon Ultra-15 centrifugal filters and quantified using RT-qPCR with primer sets targeting nucleocapsid (N1 and N2) and envelope (E) protein genes. Microbiological (faecal indicator bacteria and human and animal genetic faecal source tracking markers), epidemiological, physicochemical and hydromorphological parameters were analysed in parallel. From 44 samples, SARS-CoV-2 RNA was detected in 31, but only at 4 concentrations above the level of quantification (ranging from 8.47 × 103 to 2.07 × 104 gc/L). The results indicated that surveillance of SARS-CoV-2 RNA in surface waters as ultimate recipients could be used as an epidemiological early-warning tool in countries lacking wastewater treatment and proper sewerage infrastructure. The performance of the applied approach, including advanced sampling site characterization to trace and identify sites with significant raw sewage influence from human populations, could be further improved by adaptation of the methodology for processing higher volumes of samples and enrichment factors, which should provide the quantitative instead of qualitative data needed for WBE.

Surveillance of SARS-CoV-2 RNA in open-water sewage canals contaminated with untreated wastewater in resource-constrained regions.

Sewage-based surveillance for COVID-19 has been described in multiple countries and multiple settings. However, nearly all are based on testing sewage treatment plant inflows and outflows using structured sewage networks and treatment systems. Many resource-limited countries worldwide have open canals, lakes and other such waste-contaminated water bodies that act as a means of sewage effluent discharge. These could serve as hyperlocal testing points for detecting COVID-19 incidence using the effluents from nearby communities. However, a sensitive, robust and economical method of SARS-CoV-2 RNA detection from open waste contaminated water bodies in resource-constrained regions is currently lacking. A protocol employed in Bangalore, India, where SARS-CoV-2 RNA levels were evaluated using two open canal systems during the first and second waves in the present study. SARS-CoV-2 RNA was measured using two strategies: a modified TrueNATTM microchip-based rapid method and traditional real-time reverse transcription-PCR (rRT-PCR), which were compared for analytical sensitivity, cost and relative ease of use. SARS-CoV-2 RNA levels were detected at lower levels during the earlier half compared to the later half of the first wave in 2020. The opposite trend was seen in the second wave in 2021. Interestingly, the change in RNA levels corresponded with the community burden of COVID-19 at both sites. The modified TrueNATTM method yielded concordant results to traditional rRT-PCR in sensitivity and specificity and cost. It provides a simple, cost-effective method for detecting and estimating SARS-CoV-2 viral RNA from open-water sewage canals contaminated with human excreta and industrial waste that can be adopted in regions or countries that lack structured sewage systems.

Impact of untreated diabetes and COVID-19-related diabetes on severe COVID-19.

Diabetes is a common comorbidity in patients with coronavirus disease (COVID-19) and contributes significantly to COVID-19 severity. We aimed to investigate the association between diabetic status and severe COVID-19. This prospective study included all COVID-19 patients admitted to our hospital, who were divided into four groups according to their diabetic status: no diabetes, treated diabetes, untreated diabetes, and COVID-19-related diabetes. Severe COVID-19 was defined as a condition that required the use of a ventilator. Of the 114 patients included in this study, 26 had severe COVID-19. The adjusted odds ratio (OR; 95% confidence interval [CI]) for severe COVID-19 was significantly higher in the treated diabetes, untreated diabetes, and COVID-19-related diabetes groups than in the no diabetes group (OR: 5.9, 95% CI [1.2-27.9]; OR 12.6, 95% CI [1.8-86.4]; and OR: 9.3, 95% [1.1-81.4], respectively). Findings from this study showed that the risk of severe COVID-19 was increased in treated diabetes, untreated diabetes, and COVID-19-related diabetes compared to no diabetes. Furthermore, the OR for severe COVID-19 was greater in untreated diabetes and COVID-19-related diabetes than in treated diabetes.

A review on detection of SARS-CoV-2 RNA in wastewater in light of the current knowledge of treatment process for removal of viral fragments.

The entire globe is affected by the novel disease of coronavirus 2019 (COVID-19 or 2019-nCoV), which is formally recognised as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The World Health Organisation (WHO) announced this disease as a global pandemic. The presence of SARS-CoV-2 RNA in unprocessed wastewater has become a cause of worry due to these emerging pathogens in the process of wastewater treatment, as reported in the present study. This analysis intends to interpret the fate, environmental factors and route of transmission of SARS-CoV-2, along with its eradication by treating the wastewater for controlling and preventing its further spread. Different recovery estimations of the virus have been depicted by the detection of SARS-CoV-2 RNA in wastewater through the viral concentration techniques. Most frequently used viral concentration techniques include polyethylene glycol (PEG) precipitation, ultrafiltration, electronegative membrane, and ultracentrifugation, after which the detection and quantification of SARS-CoV-2 RNA are done in wastewater samples through quantitative reverse transcription-polymerase chain reaction (RT-qPCR). The wastewater treatment plant (WWTP) holds the key responsibility of eliminating pathogens prior to the discharge of wastewater into surface water bodies. The removal of SARS-CoV-2 RNA at the treatment stage is dependent on the operations of wastewater treatment systems during the outbreak of the virus; particularly, in the urban and extensively populated regions. Efficient primary, secondary and tertiary methods of wastewater treatment and disinfection can reduce or inactivate SARS-CoV-2 RNA before being drained out. Nonetheless, further studies regarding COVID-19-related disinfectants, environment conditions and viral concentrations in each treatment procedure, implications on the environment and regular monitoring of transmission need to be done urgently. Hence, monitoring the SARS-CoV-2 RNA in samples of wastewater under the procedure of wastewater-based epidemiology (WBE) supplement the real-time data pertaining to the investigation of the COVID-19 pandemic in the community, regional and national levels.

Detection of SARS-CoV-2 RNA in the Danube River in Serbia associated with the discharge of untreated wastewaters.

In Serbia less than 13% of collected municipal wastewaters is being treated before their release in the environment. This includes all municipal wastewater discharges from Belgrade (capital city of Serbia; population 1,700,000). Previous research has identified the impacts of raw wastewater discharges from Belgrade on the Danube River, and this study investigated if such discharges also provided a pathway for SARS-CoV-2 RNA material. Samples were collected during the most critical circumstances that occurred so far within the COVID-19 pandemics in Serbia. Grab and composite samples were collected in December 2020, during the peak of the third wave (in terms of reported cases) at the site which receives the wastewater loads in Belgrade. Grab samples collected upstream and downstream of Belgrade were also analyzed. RNA was quantified using RT-qPCR with primer sets targeting nucleocapsid (N1 and N2) and envelope (E) protein genes. SARS-CoV-2 RNA (5.97 × 103 to 1.32 × 104 copies/L) was detected only in samples collected at the site strongly impacted by the wastewaters where all three applied primer sets gave positive signals. Determined concentrations correspond to those reported in wastewater influents sampled at treatment plants in other countries indicating an epidemiological indicator function of used approach for rivers with high pollution loads in countries with poor wastewater treatment.

Comparison of virus concentration methods for the RT-qPCR-based recovery of murine hepatitis virus, a surrogate for SARS-CoV-2 from untreated wastewater.

There is currently a clear benefit for many countries to utilize wastewater-based epidemiology (WBE) as part of ongoing measures to manage the coronavirus disease 2019 (COVID-19) global pandemic. Since most wastewater virus concentration methods were developed and validated for nonenveloped viruses, it is imperative to determine the efficiency of the most commonly used methods for the enveloped severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Municipal wastewater seeded with a human coronavirus (CoV) surrogate, murine hepatitis virus (MHV), was used to test the efficiency of seven wastewater virus concentration methods: (A-C) adsorption-extraction with three different pre-treatment options, (D-E) centrifugal filter device methods with two different devices, (F) polyethylene glycol (PEG 8000) precipitation, and (G) ultracentrifugation. MHV was quantified by reverse-transcription quantitative polymerase chain reaction and the recovery efficiency was calculated for each method. The mean MHV recoveries ranged from 26.7 to 65.7%. The most efficient methods were adsorption-extraction methods with MgCl2 pre-treatment (Method C), and without pre-treatment (Method B). The third most efficient method used the Amicon® Ultra-15 centrifugal filter device (Method D) and its recovery efficiency was not statistically different from the most efficient methods. The methods with the worst recovery efficiency included the adsorption-extraction method with acidification (A), followed by PEG precipitation (F). Our results suggest that absorption-extraction methods with minimal or without pre-treatment can provide suitably rapid, cost-effective and relatively straightforward recovery of enveloped viruses in wastewater. The MHV is a promising process control for SARS-CoV-2 surveillance and can be used as a quality control measure to support community-level epidemic mitigation and risk assessment.