Evaluation of respiratory samples in etiology diagnosis and microbiome characterization by metagenomic sequencing.

BACKGROUND: The application of clinical mNGS for diagnosing respiratory infections improves etiology diagnosis, however at the same time, it brings new challenges as an unbiased sequencing method informing all identified microbiomes in the specimen. METHODS: Strategy evaluation and metagenomic analysis were performed for the mNGS data generated between March 2017 and October 2019. Diagnostic strengths of four specimen types were assessed to pinpoint the more appropriate type for mNGS diagnosis of respiratory infections. Microbiome complexity was revealed between patient cohorts and infection types. A bioinformatic pipeline resembling diagnosis results was built based upon multiple bioinformatic parameters. RESULTS: The positive predictive values (PPVs) for mNGS diagnosing of non-mycobacterium, Nontuberculous Mycobacteria (NTM), and Aspergillus were obviously higher in bronchoalveolar lavage fluid (BALF) demonstrating the potency of BALF in mNGS diagnosis. Lung tissues and sputum were acceptable for diagnosis of the Mycobacterium tuberculosis (MTB) infections. Interestingly, significant taxonomy differences were identified in sufficient BALF specimens, and unique bacteriome and virome compositions were found in the BALF specimens of tumor patients. Our pipeline showed comparative diagnostic strength with the clinical microbiological diagnosis. CONCLUSIONS: To achieve reliable mNGS diagnosis result, BALF specimens for suspicious common infections, and lung tissues and sputum for doubtful MTB infections are recommended to avoid the false results given by the complexed respiratory microbiomes. Our developed bioinformatic pipeline successful helps mNGS data interpretation and reduces manual corrections for etiology diagnosis.

AlGaN-based Schottky barrier deep ultraviolet photodetector grown on Si substrate.

This letter reports the influence of material quality and device processing on the performance of AlGaN-based Schottky barrier deep ultraviolet photodetectors grown on Si substrates. The thermal annealing can significantly improve Schottky barrier height and wet chemical etching can effectively remove etching damage. Meanwhile, the decrease of threading dislocation density and the pit size, especially the later, can substantially suppress reverse leakage. As a result, the reverse leakage current density of the as-fabricated deep UV photodetector was reduced down to 3×10-8 A/cm2. Furthermore, the responsivity of the deep UV photodetectors was greatly improved by reducing the point defect concentration.

Copper(I) Promotes Silver Sulfide Dissolution and Increases Silver Phytoavailability.

Metal sulfides, including acanthite (Ag2S), are persistent in the environment. In colloidal form, however, they can serve as a "Trojan horse", facilitating the mobility of trace metal contaminants. The natural processes that lead to the in situ dissolution of colloidal metal sulfides in soil are largely unknown. In this study, the dissolution of colloidal Ag2S in topsoil and Ag phytoavailability to wheat were examined in Ag2S-Cu(II)-thiosulfate systems. Cu(II) and thiosulfate strongly increased silver release (up to 83% of total Ag) from Ag2S in the dark. Electron paramagnetic resonance, X-ray photoelectron spectroscopy, and Cu K-edge X-ray absorption spectroscopy identified Cu(I) as the driving force of Ag2S dissolution. Density functional theory calculations further demonstrated the ability of Cu(I) to substitute for surface Ag on Ag2S in an energetically favorable manner. However, excess Cu(II) could enhance the formation of precipitates containing Cu(I), Ag, and S. Our results indicate that at ambient temperature and in the dark, Cu(I) can promote the dissolution of Ag2S and act as a precipitating agent. These findings reveal previously unrecognized biogeochemical processes of colloidal Ag2S and their importance in determining the fate of metal sulfides in the environment and probably also in vivo.

Fabrication of AlGaN nanostructures by nanolithography on ultraviolet LEDs grown on Si substrates.

AlGaN nanostructures have many applications because of their interesting and unique properties. Here we report a simple fabrication method of AlGaN nanostructures by nanolithography on ultraviolet (UV) LEDs grown on Si substrates. We also studied the effects of various AlGaN nanostructure arrays on the performance of the UV (370 nm) thin-film LEDs with an embedded n-type contact. The output power of the UV LEDs with nanostructures was enhanced by 3.9 times compared to the flat UV LEDs, while no penalty was induced for the electrical characteristics of the UV LEDs. Additionally, the far-field radiation pattern of the UV LEDs with nanostructures showed much better directionality and a much stronger intensity than the flat UV LEDs, which would be of great benefit to directional UV curing applications.

Discerning the Sources of Silver Nanoparticle in a Terrestrial Food Chain by Stable Isotope Tracer Technique.

The increasing use of silver-containing nanoparticles (NPs) in commercial products has led to NP accumulation in the environment and potentially in food webs. Identifying the uptake pathways of different chemical species of NPs, such as Ag2S-NP and metallic AgNPs, into plants is important to understanding their entry into food chains. In this study, soybean Glycine max L. was hydroponically exposed to Ag2S-NPs via their roots (10-50 mg L-1) and stable-isotope-enriched 109AgNPs via their leaves [7.9 µg (g fresh weight)-1]. Less than 29% of Ag in treated leaves (in direct contact with 109AgNP) was accumulated from root uptake of Ag2S-NPs, whereas almost all of the Ag in soybean roots and untreated leaves sourced from Ag2S-NPs. Therefore, Ag2S-NPs are phytoavailable and translocate upward. During trophic transfer the Ag isotope signature was preserved, indicating that accumulated Ag in snails most likely originated from Ag2S-NPs. On average, 78% of the Ag in the untreated leaves was assimilated by snails, reinforcing the considerable trophic availability of Ag2S-NPs via root uptake. By highlighting the importance of root uptake of Ag2S-NPs in plant uptake and trophic transfer to herbivores, our study advances current understanding of the biogeochemical fate of Ag-containing NPs in the terrestrial environment.

Different Uptake of Metal Dioxide Nanoparticles (Ceria Nanoparticles, Zirconia Nanoparticles and Silica Nanoparticles) by Wheat.

Metal dioxide nanoparticles (NPs) are produced in ever-increasing amounts and widely used in industrial, medical and consumer products. They may pose potential risks to the environment. In this study we quantitatively evaluated the phytoavailability of CeO2NPs, ZrO2NPs and SiO2NPs to wheat (Triticum aestivum L.). Metal dioxide NPs showed distinct differences in uptake, with uptake rate constants of 90.2 ± 9.9, 12.2 ± 2.5, 0.4 ± 0.02 and 0.9 ± 0.1 L kg-1 h-1 for ZrO2NPs, CeO2NPs, SiO2NPs (13 nm) and SiO2NPs (25 nm), respectively. However, such difference cannot be generalized by single factor of NP characteristics and/or root damage. This study provides fundamental information for NP uptake by crop plants.

Microbiological Diagnostic Performance of Metagenomic Next-generation Sequencing When Applied to Clinical Practice.

Background: Metagenomic next-generation sequencing (mNGS) was suggested to potentially replace traditional microbiological methodology because of its comprehensiveness. However, clinical experience with application of the test is relatively limited. Methods: From April 2017 to December 2017, 511 specimens were collected, and their retrospective diagnoses were classified into infectious disease (347 [67.9%]), noninfectious disease (119 [23.3%]), and unknown cases (45 [8.8%]). The diagnostic performance of pathogens was compared between mNGS and culture. The effect of antibiotic exposure on detection rate was also assessed. Results: The sensitivity and specificity of mNGS for diagnosing infectious disease were 50.7% and 85.7%, respectively, and these values outperformed those of culture, especially for Mycobacterium tuberculosis (odds ratio [OR], 4 [95% confidence interval {CI}, 1.7-10.8]; P < .01), viruses (mNGS only; P < .01), anaerobes (OR, ∞ [95% CI, 1.71-∞]; P < .01) and fungi (OR, 4.0 [95% CI, 1.6-10.3]; P < .01). Importantly, for mNGS-positive cases where the conventional method was inconclusive, 43 (61%) cases led to diagnosis modification, and 41 (58%) cases were not covered by empirical antibiotics. For cases where viruses were identified, broad-spectrum antibiotics were commonly administered (14/27), and 10 of 27 of these cases were suspected to be inappropriate. Interestingly, the sensitivity of mNGS was superior to that of culture (52.5% vs 34.2%; P < .01) in cases with, but not without, antibiotic exposure. Conclusions: mNGS could yield a higher sensitivity for pathogen identification and is less affected by prior antibiotic exposure, thereby emerging as a promising technology for detecting infectious diseases.

A meta-analysis of nucleos(t)ide analogues in patients with decompensated cirrhosis due to hepatitis B.

BACKGROUND: The effect of nucleos(t)ide analogues therapy in patients with decompensated cirrhosis remains unclear. AIM: The purpose of this study was to evaluate the effect of nucleos(t)ide analogues on decompensated cirrhotic patients. METHODS: An online search within PubMed, Web of Science, Embase, Cochrane Central of Register of Controlled Trials and China Biology Medicine disc from 1998-01-01 to 2011-09-05 was conducted. A meta-analysis was performed. Relative risks of mortality rate, Child-Pugh-Turcotte score and hepatitis B e-antigen (HBeAg) seroconversion of the decompensated patients were studied. RESULTS: Eight studies involving 511 patients were included. Data showed that lamivudine and telbivudine significantly decreased the mortality rate (relative risk 0.36, 95 % confidence interval 0.25-0.54), improved the Child-Pugh-Turcotte scores (mean difference -3.23, 95 % confidence interval -3.98 to -2.48) and promoted HBeAg seroconversion (relative risk 7.48, 95 % confidence interval 2.31-24.20). CONCLUSION: For patients with decompensated cirrhosis, lamivudine and telbivudine significantly decrease the mortality rate and disease severity. Also, they promote their HBeAg seroconversion.

Trophic transfer of nanomaterials and their effects on high-trophic-level predators.

Nanotechnology offers great opportunities for numerous sectors in society. One important challenge in sustainable nanotechnology is the potential of trophic transfer of nanomaterials (NMs), which may lead to unintentional impacts on environmental and human health. Here, we highlight the key advances that have been made in recent 15 years with respect to trophic transfer of heterogeneous NMs, including metal-based NMs, carbon-based NMs and nanoplastics, across various aquatic and terrestrial food chains. Particle number-based trophic transfer factors (TTFs), rather than the variable mass-based TTFs, capture the particle-specific transfer, for which NMs exhibit dynamic and complex biotransformation (e.g., dissolution, sulfidation, reduction, and corona formation). Trophic transfer of NMs has toxicological significance to predators at molecular (e.g., increased oxidative stress and modified metabolites), physiological (e.g., feeding inhibition) and population (e.g., reproduction inhibition) levels. However, linking NM exposure and toxicity remains a challenge, partly due to the dynamic biotransformation along the food chain. Although NMs have been used to increase crop yield in agriculture, they can exert detrimental impacts on crop yield and modify crop quality, depending on NMs type, exposure dose, and crop species, with unknown consequences to human health via crop consumption. Given this information, we describe the challenges and opportunities in understanding the significance of NMs trophic transfer to develop more sustainable, effective and safer nanotechnology.

Hillock Related Degradation Mechanism for AlGaN-Based UVC LEDs.

Heteroepitaxial growth of high Al-content AlGaN often results in a high density of threading dislocations and surface hexagonal hillocks, which degrade the performance and reliability of AlGaN-based UVC light emitting diodes (LEDs). In this study, the degradation mechanism and impurity/defect behavior of UVC LEDs in relation to the hexagonal hillocks have been studied in detail. It was found that the early degradation of UVC LEDs is primarily caused by electron leakage. The prominent contribution of the hillock edges to the electron leakage is unambiguously evidenced by the transmission electron microscopy measurements, time-of-flight secondary ion mass spectrometry, and conductive atomic force microscopy. Dislocations bunching and segregation of impurities, including C, O, and Si, at the hillock edges are clearly observed, which facilitate the trap-assisted carrier tunneling in the multiple quantum wells and subsequent recombination in the p-AlGaN. This work sheds light on one possible degradation mechanism of AlGaN-based UVC LEDs.

Key knowledge gaps for One Health approach to mitigate nanoplastic risks.

There are increasing concerns over the threat of nanoplastics to environmental and human health. However, multidisciplinary barriers persist between the communities assessing the risks to environmental and human health. As a result, the hazards and risks of nanoplastics remain uncertain. Here, we identify key knowledge gaps by evaluating the exposure of nanoplastics in the environment, assessing their bio-nano interactions, and examining their potential risks to humans and the environment. We suggest considering nanoplastics a complex and dynamic mixture of polymers, additives, and contaminants, with interconnected risks to environmental and human health. We call for comprehensive integration of One Health approach to produce robust multidisciplinary evidence to nanoplastics threats at the planetary level. Although there are many challenges, this holistic approach incorporates the relevance of environmental exposure and multi-sectoral responses, which provide the opportunity to identify the risk mitigation strategies of nanoplastics to build resilient health systems.

Effect of metal cations on antimicrobial activity and compartmentalization of silver in Shewanella oneidensis MR-1 upon exposure to silver ions.

Silver is an antimicrobial agent that is used extensively in consumer products, such as fabrics and humidifiers. Silver ion (Ag+) uptake in bacteria represents a crucial phase of antimicrobial activity. However, the uptake mechanism of Ag+ in bacteria remains largely unknown. The genus Shewanella drives many geochemical processes of nutrients and pollutants in soils. In the present study, Ag+ uptake by Shewanella oneidensis MR-1 was first investigated in a laboratory in defined anaerobic, oligotrophic, and inorganic media with or without cations (potassium ions [K+], magnesium ions [Mg2+], and zinc ions [Zn2+]). Our results revealed variations in antimicrobial activity of Ag+ in the presence of Mg2+ and Zn2+. First, Mg2+ significantly decreased antimicrobial activity of Ag+ in S. oneidensis MR-1 by inhibiting cellular Ag+ uptake when compared with K+. The results were consistent with that of Co2+ (Mg2+ channel blocker) decreased Ag+ uptake by S. oneidensis MR-1. Moreover, Mg2+ promoted riboflavin secretion and facilitated the formation of metallic Ag nanoparticles on bacterial surfaces, which was beneficial for extracellular electron transfer and consequently reduced antibacterial activity of Ag+. Second, Zn2+ increased the antimicrobial activity of Ag+ in S. oneidensis MR-1, although the effect on Ag+ uptake was minimal. A synergistic interaction between Zn2+ and Ag+ led to an increase in dead cells and decreased ferrihydrite reduction capacity. The findings suggest that Mg2+ could reduce the environmental risk of Ag+ to soil bacteria, while Zn2+ should be of particular concern due to its synergistic antimicrobial effect on bacteria.

A biological and genomic comparison of a drug-resistant and a drug-susceptible strain of Candida auris isolated from Beijing, China.

The fungal pathogen Candida auris has emerged as a new threat to human health. We previously reported the first isolate of C. auris (BJCA001) in China, which belongs to the South Asian clade (I) and was susceptible to all antifungals tested. In this study, we report the isolation of a drug-resistant C. auris strain (BJCA002) from the same city (Beijing). Strain BJCA002 belongs to the South African clade (III) and is resistant to fluconazole and amphotericin B based on the tentative MIC breakpoints. Taking advantage of the two isolates with distinct antifungal susceptibility and genetic origins, we performed a biological and genomic comparative study. Besides antifungal susceptibility, strains BJCA001 and BJCA002 showed differences in multiple aspects including morphologies, expression of virulence factors, virulence, mating type, and genomic sequence and organization. Notably, strain BJCA002 was less virulent than BJCA001 in both the Galleria mellonella and mouse systemic infection models. Genomic analysis demonstrated that strain BJCA002 but not BJCA001 had multiple mutations in drug resistance-associated genes, including a hot-spot mutation of ERG11 (VF125AL, namely V125A and F126L) and some missense mutations in CDR1, MDR1, and TAC1. Notably, strain BJCA001 carried 64 copies of the Zorro3 retrotransposon, whereas BJCA002 had only 3 copies in the genome. Taken together, our findings not only reveal the genetic and phenotypic diversities of the two isolates from Beijing, China, but also shed new light on the genetic basis of the antifungal resistance and virulence of C. auris.

A rare case of adrenal gland abscess due to anaerobes detected by metagenomic next-generation sequencing.

A 50-year-old woman presented with right back pain, low fever, leukocytosis, a high level of C-reactive protein and a high erythrocyte sedimentation rate. Abdominal magnetic resonance imaging (MRI) revealed a hypodense lesion in the right suprarenal region, while PET/CT showed mildly increased metabolic activity. A CT-guided percutaneous puncture was performed, and foul-smelling thick pus was removed, which indicated an anaerobic infection. A smear of the pus showed both gram-positive and gram-negative microorganisms. Traditional culture only detected Escherichia coli, Proteus mirabilis, and Actinomyces turicensis. While surprisingly, metagenomic next-generation sequencing (mNGS) of both the pus and blood showed high reads of multiple pathogens, including anaerobes and the three culture-positive pathogens. Thus, adrenal gland abscess was confirmed, and a combination therapy of catheter drainage and effective antimicrobial treatment was started. Six days later, the patient had clinically improved and mNGS showed dramatically decreased reads of all pathogens. A follow-up lab examination of inflammatory biomarkers was normal, and the adrenal mass was reduced radiographically.

Environmental and human health risks from metal exposures nearby a Pb-Zn-Ag mine, China.

Metal contamination in mining areas, where mining and farming coexist, is of great concern worldwide. Nevertheless, a disconnection exists between those risks to environmental and human health. In this study, an integrated approach was used to connect the environmental and human health risks from metal exposures nearby a Pb-Zn-Ag mine. The field survey showed metal contamination in soils and crop plants as well as variation in soil microbial community in mining region relative to the reference site. Together with non-carcinogenic and carcinogenic risk assessment with a probabilistic approach, Cd and Cr were identified as the priority contaminants. Further, consumption of contaminated food crops was a significant pathway of human exposure. Especially, children were susceptible to metal contamination with non-carcinogenic hazard index (0.45 for soil ingestion and 1.19 for dietary intake) and carcinogenic risk index (3.47 × 10-4 for soil ingestion and 5.10 × 10-3 for dietary intake) at the 50th percentile. These findings facilitate the priority actions on mitigation strategies to minimize the environmental and health risks. Also, the potential environmental and human health consequences due to uncontrolled mining in this region serve as a case study for other regions involved in mining activities.

Diagnostic accuracy of metagenomic next-generation sequencing for active tuberculosis in clinical practice at a tertiary general hospital.

BACKGROUND: To evaluate the diagnostic accuracy of metagenomic next-generation sequencing (mNGS) for active tuberculosis (TB). METHODS: We retrospectively collected 820 samples at Zhongshan Hospital, Fudan University in Shanghai, China, between 1 April 2017 and 31 March 2018. They were classified into TB cases (125, 15.2%) and NOT TB cases (695, 84.8%) according to the clinical diagnosis. Specimens were evaluated by a regular clinical microbiological assay and mNGS performed in parallel. RESULTS: Sixty-one confirmed TB cases and 64 clinical TB cases were included. The overall sensitivity of mNGS was 49.6% [95% confidence interval (95% CI), 40.6-58.6%], and the specificity was 98.3% (95% CI, 96.9-99.1%), with peak sensitivities of 88.9% (95% CI, 50.7-99.4%) for lung tissue, 55.0% (95% CI, 32.0-76.2%) for bronchoalveolar lavage fluid (BALF), and 50.0% (95% CI, 32.8-67.2%) for serous fluids. The overall sensitivity of mNGS was superior to that of the culture assay (35.2%, 95% CI, 27.0-44.3), but no superior sensitivity for sputum was observed in mNGS compared with the culture assay (mNGS: 52.3%, 95% CI, 31.1-72.6%; culture: 60.9%, 95% CI, 38.8-79.5%). In clinical TB cases, mNGS detected additional positive results (40.6%, 26/64). mNGS reduced the turnaround time from 2-6 weeks to 32-36 hours. CONCLUSIONS: mNGS may be a promising technology for the early auxiliary diagnosis of active TB, especially sputum-negative pulmonary TB (PTB) and tuberculous serous effusion.

The clinical value of metagenomic next-generation sequencing in the microbiological diagnosis of skin and soft tissue infections.

BACKGROUND: Metagenomic next-generation sequencing (mNGS), with its comprehensiveness, is widely applied in microbiological diagnosis. Etiological diagnosis is of paramount clinical importance in patients with skin and soft tissue infections (SSTIs). However, the clinical application of mNGS in SSTIs is relatively less studied. MATERIALS AND METHODS: From April 1, 2017 to December 31, 2019, 96 SSTI cases were collected. The positive rates of pathogens detected by mNGS and culture were compared by analyzing tissue samples, pus, swabs, and/or interstitial fluids obtained from the infected parts. Modification of the antibiotic treatment strategy due to mNGS was also assessed. RESULTS: The sensitivity of mNGS for detecting pathogens in SSTI cases was superior to that of culture testing (67.7% vs 35.4%; p < 0.01). Significantly higher identification rates for viruses (10.4% vs 0.0%; p < 0.01) and anaerobes (11.5% vs 1.0%; p < 0.01) were obtained with mNGS compared to culture. Of note, rare pathogens such as Vibrio vulnificus and Bartonella henselae were also detected by mNGS. Importantly, the proportion of multi-pathogen SSTIs detected by mNGS was higher than that of multi-pathogen SSTIs detected by culture (16.7% vs 6.3%; p = 0.035). The rate of targeted antibiotic treatment was significantly higher in mNGS-positive cases than in mNGS-negative cases (41.7% vs 3.8%; p < 0.01). In culture-negative and mNGS-positive cases, the improvement rate was higher than that in mNGS-negative cases, but this was not statistically significant (75.0% vs 73.1%; p = 0.864). CONCLUSIONS: mNGS is a promising tool for the etiological diagnosis of SSTIs, particularly in identifying viruses, anaerobes, and multi-pathogen infections. The application of mNGS testing in clinical practice could change antibiotic treatment strategies and partly benefit clinical outcomes.

Arthritis caused by Legionella micdadei and Staphylococcus aureus: metagenomic next-generation sequencing provides a rapid and accurate access to diagnosis and surveillance.

Legionella spp. is an important pulmonary pathogen but rarely causes extra-pulmonary infections. We report a case of joint infection caused by Legionella micdadei and Staphylococcus aureus in a 54-year-old male with medication history of oral steroid for systemic lupus erythematosus (SLE). He developed arthritis in his right metacarpophalangeal (MCP) joints without precursor pneumonia. In the joint aspirate, S. aureus was detected through culture. The existence of L. micdadei and S. aureus were indicated by metagenomic next-generation sequencing (mNGS) and confirmed by 16S rRNA sequence analysis. After oral levofloxacin treatment for 54 days, the patient's symptoms ameliorated and blood test results improved, which were consistent with the dynamic trend of reads numbers in mNGS data. Our case included, arthritis caused by Legionella spp. have been reported in 11 patients. However, our case is the first to report septic arthritis caused by L. micdadei in native joints and monitored by mNGS. This case demonstrated an application of mNGS for etiological diagnosis and semi-quantification in joint aspirate. mNGS may serve as a promising tool for rapid and accurate etiological diagnosis and surveillance, contributing to appropriate antimicrobial drug applications and timely medication adjustments when necessary.

Significant contribution of metastable particulate organic matter to natural formation of silver nanoparticles in soils.

Particulate organic matter (POM) is distributed worldwide in high abundance. Although insoluble, it could serve as a redox mediator for microbial reductive dehalogenation and mineral transformation. Quantitative information on the role of POM in the natural occurrence of silver nanoparticles (AgNPs) is lacking, but is needed to re-evaluate the sources of AgNPs in soils, which are commonly considered to derive from anthropogenic inputs. Here we demonstrate that POM reduces silver ions to AgNPs under solar irradiation, by producing superoxide radicals from phenol-like groups. The contribution of POM to the naturally occurring AgNPs is estimated to be 11-31%. By providing fresh insight into the sources of AgNPs in soils, our study facilitates unbiased assessments of the fate and impacts of anthropogenic AgNPs. Moreover, the reducing role of POM is likely widespread within surface environments and is expected to significantly influence the biogeochemical cycling of Ag and other contaminants that are reactive towards phenol-like groups.

Screening of wheat straw biochars for the remediation of soils polluted with Zn (II) and Cd (II).

The immobilization behaviors of Zn(II) and Cd(II) by wheat straw (WS) biochars could vary with the soil conditions. In the acidic environment, WS biochars produced at low temperature were more competent than those produced at high temperature on Zn(II) and Cd(II) immobilization; while WS biochars produced at high temperature were more effective than those produced at low temperature in the alkaline environment. The ions in the porewater could compromise the sorption capacities of Zn(II) and Cd(II) by WS biochars in acidic soils, while could enhance them in alkaline soils. For biochars produced at the same temperature, residence time had little effect on their behaviors of Zn(II) and Cd(II) immobilization. Only a small portion of immobilized Zn(II)/Cd(II) could be released from WS biochar in the simulated acid rain. Compared with Zn(II)/Cd(II) adsorbed on the acidic functional groups, Zn(II)/Cd(II) precipitates were more stable in 0.01 M CaCl2 solution. Most of the Zn(II) and Cd(II) species on biochar could be released in 1 mM citric acid solution. The immobilized Zn(II) and Cd(II) on WS biochar are likely to be released into the soil environment in the long run.