Environmental surveillance of SARS-CoV-2 RNA in wastewater systems and related environments in Wuhan: April to May of 2020.

Wastewater-based epidemiology (WBE) has emerged as an effective environmental surveillance tool in monitoring fecal-oral pathogen infections within a community. Congruently, SARS-CoV-2, the etiologic agent of COVID-19, has been demonstrated to infect the gastrointestinal tissues, and be shed in feces. In the present study, SARS-CoV-2 RNA was concentrated from wastewater, sludge, surface water, ground water, sediment, and soil samples of municipal and hospital wastewater systems and related environments in Wuhan during the COVID-19 middle and low risk periods, and the viral RNA copies quantified using reverse transcription quantitative polymerase chain reaction (RT-qPCR). From the findings of this study, during the middle risk period, one influent sample and three secondary effluents collected from waste water treatment plant 2, as well as two samples from Jinyintan Hospital wastewater system influent were SARS-CoV-2 RNA positive. One sludge sample collected from Guanggu Branch of Tongji Hospital, which was obtained during the low risk period, was also positive for SARS-CoV-2 RNA. These study findings demonstrate the significance of WBE in continuous surveillance of SARS-CoV-2 at the community level, even when the COVID-19 prevalence is low. Overall, this study can be used as an important reference for contingency management of wastewater treatment plants and COVID-19 prevention and control departments of Wuhan.

Integrating MFT-qPCR techniques in constructed wetland faecal bacterial purification monitoring; a case of a typical tropical hybrid constructed wetland system.

The sanitation control of pathogens in the tropical effluents needs much more attention to ensure ecosystem health integrity and the safety of human health. The common use of chemicals in achieving this in wastewater treatment has remained unsustainable due to much health concern. Indeed, based on the numerous challenges associated with faecal pathogenic bacteria in wastewaters, the focus is now on achieving higher purification efficiencies in the elimination of the human pathogens from wastewater through eco-sustainable systems such as constructed wetlands (CWs). Hence, the need to explore the application of constructed wetlands in wastewater treatment under specific local environmental conditions for accurate understanding and improved treatment efficiency. This study therefore aimed at monitoring constructed wetlands faecal bacteria purification efficiency through integrated non-molecular membrane filtration technique and molecular quantitative polymerase chain reaction (MFT-qPCR) technique. The results showed some shortfall in the treatment system and also proved that integrating MFT-qPCR in faecal bacterial purification monitoring within a constructed wetland system provides a more accurate and reliable outcome. Additionally, the wetland purification efficiency was low (<80%) with the dissolved oxygen posing the strongest influence on faecal pathogenic bacterial purification trend across the wetland. Hence, the need to regularly carry out dredging and macrophyte harvesting as well as the use of holistic and more integrative approaches such as MFT-qPCR in managing and monitoring the performance of CWs in faecal pathogen eradication for improved CWs purification efficiency.

COVID-19 pandemic: Water, sanitation and hygiene (WASH) as a critical control measure remains a major challenge in low-income countries.

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is responsible for the deadly respiratory disease called coronavirus disease of 2019 (COVID-19), an ongoing global public health emergency that has been declared a pandemic by the World Health Organization. We review literature on the transmission and control of SARS-CoV-2 and discuss the challenges of focusing on water, sanitation and hygiene (WASH) as critical control measures in low-income countries. A significantly higher prevalence of SARS-CoV-2 infection and COVID-19 related deaths has been reported for the United States of America and other high-income countries in Europe and Asia, regardless of advanced medical facilities in those countries. In contrast, much lower COVID-19 related morbidity and mortality rates have been documented in many low-income countries, despite having comparatively higher socioeconomic burdens and suboptimal medical facilities. By September 29, 2020 over one million deaths have been reported. On the same day, the cumulative total of COVID-19 related morbidity for Africa was 35,954 with 3.5% of the global COVID-19 related deaths. We present arguments for the relatively low COVID-19 morbidity and mortality rates in many low-income countries and discuss the critical importance of WASH for preventing the spread of infectious diseases like COVID-19. We observe that the key recommendations put forward by the World Health Organization to effectively control the pandemic have been difficult to implement in low-income countries. We conclude that the pandemic reinforces previous pronouncements that adequate and effective WASH measures are crucial for public health and recommend closer coordination between public health and WASH sectors.

Effects of microcystin-producing and non-microcystin-producing Microcystis on the behavior and life history traits of Chironomus pallidivittatus.

Species of the genus Microcystis are among the most notorious cyanobacteria in eutrophic lakes worldwide, with ability present adverse effects on many aquatic organisms. In the surface sediments, Microcystis can be ingested by benthic macroinvertebrates such as Chironomus. However, the potential negative effects of Microcystis on Chironomus life history traits remain unclear. In the present study, we investigated the effect of different Microcystis diets on specific behaviors (burrowing activity, locomotion ability) and life history traits of Chironomus pallidivittatus (Diptera, Chironomidae). We also studied the interactive effects of microcystin-producing M. aeruginosa and temperature (15, 20, and 25 °C) stress on chironomid larvae. The results showed that the inhibitory effect on the cumulative emergence and burrowing activity of larvae was more severe when they were fed M. aeruginosa among the three Microcystis diets groups. Locomotion ability (i.e., locomotor distance and velocity) and adult dry weight decreased significantly in the group fed M. aeruginosa. Locomotion was significantly inhibited and mortality increased when the larvae were fed a mixture of M. aeruginosa and M. wesenbergii, which may have been the result of additive or synergistic effect of the toxins. Under the stress of lower temperature, C. pallidivittatus larvae exhibited weaker locomotion and growth ability, and the emerging adults were mostly male. At both the lower and higher temperature conditions, M. aeruginosa cause cumulative emergence decreased, and sex ratio imbalance, which inhibited the reproduction of larvae from the population perspective. The fourth-instar larvae showed better adaption to Microcystis than did the other instars. This study thus highlights the adverse effects of microcystin-producing M. aeruginosa on Chironomus. It also provides a novel perspective on how environmental factors may influence the behavior and life history traits of chironomid larvae, and how they may respond to cyanobacterial blooms and global warming.

Bioflocculation effect of Glyptotendipes tokunagai on different Microcystis species: Interactions between secreted silk and extracellular polymeric substances.

Cyanobacterial blooms are a major problem in many lakes and can negatively impact public health and ecosystem services. The bioflocculation technique has proven to be a cost-effective, environmentally friendly technique with no secondary pollution to harvest multiple microalgae; however, few studies have focused on its effect on and potential for controlling cyanobacterial blooms in eutrophic lakes. In this study, the bioflocculation efficiencies of different Microcystis species under Glyptotendipes tokunagai (Diptera, Chironomidae) stress conditions and the interactions between secreted silk from Chironomid larvae and extracellular polymeric substances (EPS) from Microcystis were compared. The results indicated that G. tokunagai presented better bioflocculation efficiency on M. wesenbergii than on M. aeruginosa. The formation of "Large Algal Aggregate" flocs was promoted by the derived-soluble extracellular polymeric substances (i.e., proteins and polysaccharides, sEPS) from M. wesenbergii and silk from G. tokunagai. Both M. wesenbergii and midge silk had abundant functional groups, which was beneficial to the formation of the large aggregate. G. tokunagai secreted a large amount of silk to bridge with the sEPS of M. wesenbergii, forming a network structure via interaction between filamentous substance (i.e., complex of sEPS and silk) that plays an important role in the aggregation of Microcystis and the removal of the Microcystis biomass in the water column. The findings provide further insights that will benefit the existing efforts of combating Microcystis blooms in the water column via bioflocculation and will provide a new sustainable approach for inhibiting early bloom formation from the perspective of its provenance in the sediment-water interface.

Co-regulatory role of Microcystis colony cell volume and compactness in buoyancy during the growth stage.

The buoyancy of Microcystis colonies determines the occurrence and dominance of bloom on the water surface. Besides the cell density regulation and the formation of larger size aggregates, increases in cell volume per colony (Vcell) and the colony's compactness (i.e., volume ratio of cells to the colony, VR) may promote Microcystis colony buoyancy. Yet only a few studies have studied the relationship between the internal structure variation of colonies and their buoyancy, and the co-regulatory role of Vcell and VR of Microcystis colonies in the floating velocity (FV) remains largely unexplored. In the present study, we optimized a method for measuring the compactness of Microcystis colonies based on the linear relationship between total Vcell and chlorophyll a. Different relationships between the VRs and FVs were observed with different colony size and Vcell range groups. Both field and laboratory experiments showed that FV/(D50, median diameter)2 had a significant linear relationship with VR, indicating that the cell density and extracellular polysaccharides were unchanged over a short time period and could be estimated via the slope and intercept of a fitted line. We also constructed a functional relationship between FV, VR, and Vcell and found that high VR and Vcell can promote Microcystis buoyancy. This means that increasing cell compactness or Vcell may be an active regulation strategy for Microcystis colonies to promote buoyancy. Therefore, quantifying the internal structure of Microcystis colonies is strongly recommended for the assessment of Microcystis bloom development and their management. Graphical abstract.

Denitrification characterization of dissolved oxygen microprofiles in lake surface sediment through analyzing abundance, expression, community composition and enzymatic activities of denitrifier functional genes.

The responses of denitrifiers and denitrification ability to dissolved oxygen (DO) concent in different layers of surface lake sediments are still poorly understood. Here, the optimal denitrification condition was constructed based on response surface methodology (RSM) to analyze the denitrification characteristics of surface sediments. The aerobic zone (AEZ), hypoxic zone (HYZ), up-anoxic zone (ANZ-1) and sub-anoxic zone (ANZ-2) were partitioned based on the oxygen contents, and sediments were collected using a customized-designed sub-millimeter scale sampling device. Integrated real-time quantitative PCR, Illumina Miseq-based sequencing and denitrifying enzyme activities analysis revealed that denitrification characteristics varied among different DO layers. Among the four layers, the DNA abundance and RNA expression levels of norB, nirS and nosZ were the highest at the aerobic layer, hypoxic layer and up-axoic layer, respectively. The hypoxia and up-anaerobic layer were the active nitrogen removal layers, since these two layers displayed the highest DNA abundance, RNA expression level and enzyme activities of denitrification functional genes. The abundance of major denitrifying bacteria showed significant differences among layers, with Azoarcus, Pseudogulbenkiania and Rhizobium identified as the main nirS, nirK and nosZ-based denitrifiers. Pearson's correlation revealed that the response of denitrifiers to environmental factors differed greatly among DO layers. Furthermore, napA showed higher DNA abundance and RNA expression level in the aerobic and hypoxic layers than anaerobic layers, indicating that aerobic denitrifiers might play important roles at these layers.

Efficacy of zero nitrous oxide emitting aerobic denitrifying bacterium, Methylobacterium gregans DC-1 in nitrate removal with strong auto-aggregation property.

A novel aerobic denitrifying strain Methylobacterium gregans DC-1 was isolated and identified. Strain DC-1 removed 98.4% of nitrate-nitrogen (NO3--N) and 80.7% of total organic carbon with initial concentrations of 50 and 2400 mg/l, respectively. The N balance showed that most NO3--N was converted to N2 (62.18%) without nitrous oxide (N2O) emission. Response surface analysis showed that the optimal conditions for total N removal were carbon (C):N ratio of 18.7, temperature of 26.8 °C, pH of 6.5 and shaking speed of 180 rpm. In combination with the N balance and successful amplification of napA, nirK and nosZ genes, the metabolic pathway was as follows: NO3-NO2- → NO → N2O → N2. Strain DC-1 had strong auto-aggregation rate (maximum 38.7%), produced large amounts of extracellular polymeric substances (EPS; maximum of 781.4 mg/g cell dry weight) and had corresponding strong hydrophobicity (maximum 83.2%). Pearson correlation analysis showed that EPS content and hydrophobicity were significantly positively correlated with auto-aggregation.

Comparison of intestinal microbes in female and male Chinese concave-eared frogs (Odorrana tormota) and effect of nematode infection on gut bacterial communities.

The Chinese concave-eared frog (Odorrana tormota) is a rare and threatened species with remarkable sexual dimorphism. Intestinal microbes are understood to play important roles in animal physiology, growth, ecology, and evolution. However, little is known about the intestinal microbes in female and male frogs, as well as the contributing effect by gut infesting nematodes to the co-habiting bacteria and their function in degradation food rich in chitin. Here, this study analyzed the microbiota of the intestinal tract of both female and male, healthy as well as nematode-infested concave-eared frogs using high throughput 16S rRNA sequencing and metagenomic techniques. The results showed that the bacterial composition of the microbiota at the phylum level was dominated by Firmicutes, Verrucomicrobia, Bacteroidetes, and Proteobacteria. The study also revealed that the community composition below the class level could be represent sex differences, particularly with regard to Enterobacteriales, Enterobacteriaceae, Peptostreptococcaceae, and Rikenellaceae, among others. Carbohydrate-active enzyme-encoding genes and modules were identified in related gut bacteria by metagenomic analysis, with Bacteroidia, Clostridia, and gammaproteobacteria predicted to be the main classes of chitin-decomposing bacteria in the frog intestine. In addition, the abundance of some bacteria significantly increased or decreased in nematode-infected hosts compared with healthy individuals, including Verrucomicrobia, Verrucomicrobiae, Negativicutes, Actinobacteria, and Bacilli, among others. This indicates that nematode infection may affect the richness and composition of some gut bacteria.

Novel heterotrophic nitrogen removal and assimilation characteristic of the newly isolated bacterium Pseudomonas stutzeri AD-1.

AD-1, an aerobic denitrifier, was isolated from activated sludge and identified as Pseudomonas stutzeri. AD-1 completely removed NO3- or NO2- and removed 99.5% of NH4+ during individual culturing in a broth medium with an initial nitrogen concentration of approximately 50 mg L-1. Results showed that larger amounts of nitrogen were removed through assimilation by the bacteria. And when NH4+ was used as the sole nitrogen source in the culture medium, neither NO2- nor NO3- was detected, thus indicating that AD-1 may not be a heterotrophic nitrifier. Only trace amount of N2O was detected during the denitrification process. Single factor experiments indicated that the optimal culture conditions for AD-1 were: a carbon-nitrogen ratio (C/N) of 15, a temperature of 25°C and sodium succinate or glucose as a carbon source. In conclusion, due to the ability of AD-1 to utilize nitrogen of different forms with high efficiencies for its growth while producing only trace emissions of N2O, the bacterium had outstanding potential to use in the bioremediation of high-nitrogen-containing wastewaters. Meanwhile, it may also be a proper candidate for biotreatment of high concentration organic wastewater.