Revealing the comprehensive impact of organic compounds on the partial nitrification-anammox system during incineration leachate treatment: metabolic hierarchy and adaptation.

Organics, as widespread pollutants in high-strength ammonia wastewater, typically exert adverse effects on the performance of partial nitrification-anammox (PNA) systems. However, the in-depth knowledge on how microbial consortia respond to these disturbances remains limited. In this study, we unveiled the evolution of complex organic matter flow and its impact on the metabolic hierarchy and adaptation of microbial consortia, employing multi-omics approaches, i.e., 16S amplicon sequencing, metagenomics, and metabolomics. In a two-stage PNA system sequentially treating synthetic wastewater and incineration leachate over 230 days, partial nitrification stayed stable (nitrite accumulation > 97%) while anammox efficiency dropped (nitrogen removal decreased from 86% to 78%). The phenomenon was revealed to be correlated with the evolution of dissolved organic matter (DOM) and xenobiotic organic compounds (XOCs). In the PN stage, ammonia-oxidizing bacteria (AOB) exhibited excellent adaptability through active metabolic regulation after treating leachate. Numerous heterotrophs proliferated to utilize DOM and XOCs, triggering a "boom" state evident in the glycerophospholipid metabolism. However, in the anammox stage, the competition between carbon fixation and central carbon metabolism within autotrophs and heterotrophs became evident. Increased biosynthesis costs inhibited the central metabolism (specific anammox activity decreased by 66%) and the Wood-Ljungdahl pathway of anammox bacteria (AnAOB) in the presence of recalcitrant organics. Additionally, the degradation of organics was limited, exhibiting a "bust" state. This study revealed the metabolic adaption and susceptibility of AOB and AnAOB in response to organics from the leachate, demonstrating the applicability of the two-stage configuration for treating high-strength wastewater containing abundant and diverse organics.

Innovative determination of the specific anammox activity for anammox sludge from continuous flow reactors: A comparison between continuous flow test and batch test.

A novel method for measuring specific anammox activity (SAA) was proposed based on continuous flow tests to accurately determine the SAA of anammox sludge from continuous flow reactors, resolving the challenges of inaccurate SAA assessment caused by substrate shock to anammox bacteria. Results showed SAA of expanded granular sludge bed sludge via batch tests (0.101 ± 0.018 g-N·g-VSS-1·d-1) was lower than continuous flow tests (0.206 ± 0.010 g-N·g-VSS-1·d-1) (p < 0.05), highlighting the impact of substrate shock. Conversely, SAA of sequencing batch reactor sludge assessed via batch tests (0.878 ± 0.008 g-N·g-VSS-1·d-1) was higher than continuous flow tests (0.809 ± 0.005 g-N·g-VSS-1·d-1) (p < 0.01), attributed to endogenous denitrification. The advantages of continuous flow tests over batch tests included milder feeding way, stricter anaerobic conditions, and minimal sampling impact on system. Our study contributes to more accurate measurements of SAA of anammox sludge from continuous flow reactors, favoring long-term robust operation of anammox reactors.

Metagenomics insight into the long-term effect of ferrous ions on the mainstream anammox system.

Anaerobic ammonium oxidation (anammox) bacteria have a high requirement for iron for their growth and metabolism. However, it remains unclear whether iron supplementation can sustain the stability of mainstream anammox systems at varying temperatures. Here, we investigated the long-term effects of Fe2+ on the mainstream anammox systems. Our findings revealed that the nitrogen removal efficiency (NRE) of the anammox system supplemented with 5 mg/L Fe2+ decreased from 76.5 ± 0.76% at 35 °C to 39.0 ± 9.9% at 25 °C. Notably, higher dosages of Fe2+ (15 mg/L and 30 mg/L) inhibited the anammox system, resulting in NREs of 15.9 ± 8.1% and 2.5 ± 1.1% at 25 °C, respectively. The results of microbial communities and function profiles suggested that the high Fe2+ dosage seriously affected the iron assimilation and utilization in the mainstream anammox system. This was evident from the decreased abundance of genes associated with Fe(II) transport and uptake, which in turn hindered the biosynthesis of intracellular iron-cofactors, resulting in decrease in the absolute abundance of Candidatus Brocadia, a key anammox bacterium, as well as a decline in NRE. Furthermore, our results showed that the anammox process was more susceptible to iron supplementation at 25 °C compared to 35 °C, which may be due to the oxidative stress reactions induced by combined lowered temperature and a high Fe2+ dosage. Overall, these findings offer a deeper understanding of the effect of iron in mainstream anammox systems, which can contribute to improved stability maintenance and effectiveness of anammox processes.

Sulfur autotrophic denitrification as an efficient nitrogen removals method for wastewater treatment towards lower organic requirement: A review.

The sulfur autotrophic denitrification (SADN) process is an organic-free denitrification process that utilizes reduced inorganic sulfur compounds (RISCs) as the electron donor for nitrate reduction. It has been proven to be a cost-effective and environment-friendly approach to achieving carbon neutrality in wastewater treatment plants. However, there is no consensus on whether SADN can become a dominant denitrification process to treat domestic wastewater or industrial wastewater if organic carbon is desired to be saved. Through a comprehensive summary of the SADN process and extensive discussion of state-of-the-art SADN-based technologies, this review provides a systematic overview of the potential of the SADN process as a sustainable alternative for the heterotrophic denitrification (HD) process (organic carbons as electron donor). First, we introduce the mechanism of the SADN process that is different from the HD process, including its transformation pathways based on different RISCs as well as functional bacteria and key enzymes. The SADN process has unique theoretical advantages (e.g., economy and carbon-free, less greenhouse gas emissions, and a great potential for coupling with novel autotrophic processes), even if there are still some potential issues (e.g., S intermediates undesired production, and relatively slow growth rate of sulfur-oxidizing bacteria [SOB]) for wastewater treatment. Then we present the current representative SADN-based technologies, and propose the outlooks for future research in regards to SADN process, including implement of coupling of SADN with other nitrogen removal processes (e.g., HD, and sulfate-dependent anaerobic ammonium oxidation), and formation of SOB-enriched biofilm. This review will provide guidance for the future applications of the SADN process to ensure a robust-performance and chemical-saving denitrification for wastewater treatment.

Oxidative stress and antioxidant mechanisms of obligate anaerobes involved in biological waste treatment processes: A review.

In-depth understanding of the molecular mechanisms and physiological consequences of oxidative stress is still limited for anaerobes. Anaerobic biotechnology has become widely accepted by the wastewater/sludge industry as a better alternative to more conventional but costly aerobic processes. However, the functional anaerobic microorganisms used in anaerobic biotechnology are frequently hampered by reactive oxygen/nitrogen species (ROS/RNS)-mediated oxidative stress caused by exposure to stressful factors (e.g., oxygen and heavy metals), which negatively impact treatment performance. Thus, identifying stressful factors and understanding antioxidative defense mechanisms of functional obligate anaerobes are crucial for the optimization of anaerobic bioprocesses. Herein, we present a comprehensive overview of oxidative stress and antioxidant mechanisms of obligate anaerobes involved in anaerobic bioprocesses; as examples, we focus on anaerobic ammonium oxidation bacteria and methanogenic archaea. We summarize the primary stress factors in anaerobic bioprocesses and the cellular antioxidant defense systems of functional anaerobes, a consortia of enzymatic and nonenzymatic mechanisms. The dual role of ROS/RNS in cellular processes is elaborated; at low concentrations, they have vital cell signaling functions, but at high concentrations, they cause oxidative damage. Finally, we highlight gaps in knowledge and future work to uncover antioxidant and damage repair mechanisms in obligate anaerobes. This review provides in-depth insights and guidance for future research on oxidative stress of obligate anaerobes to boost the accurate regulation of anaerobic bioprocesses in challenging and changing operating conditions.

A versatile control strategy based on organic carbon flow analysis for effective treatment of incineration leachate using an anammox-based process.

Anammox-based process provides an alternative for the sustainable treatment of incineration leachate that has high-load ammonium and high residual heat, but the high concentrations of organics in such leachates brought challenges for the process control. For the first time, a two-stage partial nitrification (PN)-anammox process coupled with a pre-enhanced anaerobic digestion (AD) was established to achieve efficient nitrogen removal from incineration leachate. Satisfactory nitrogen and chemical oxygen demand (COD) removal efficiencies were achieved-with the average values of 90% and 78%, respectively-despite fluctuating influent properties [1100-2000 mg-total nitrogen (TN)/L and 3800-15800 mg-COD/L]. A versatile control strategy was developed to create an optimum autotrophic environment for nitrifier and anammox bacteria: i) enhanced AD set before the PN-anammox process captured nearly 50% of the influent COD; ii) in the PN unit, ammonia-oxidizing bacteria were well adapted to COD concentrations of 1420-2400 mg/L, and dissolved oxygen (0.2-0.4 mg/L) controlling combined with a high free nitrous acid concentration (>0.08 mg/L) ensured a nitrite accumulation rate of >95%; and iii) in the anammox unit, a suitable influent NO2--N/NH4+-N ratio (the average value of 1.27) was achieved by mixing AD effluent with PN effluent (1:1.78, v/v), contributing to a high TN removal of 78 ± 2.4%. Nevertheless, 980-1560 mg/L of COD remained in the influent of the anammox unit; biorefractory humic acids in this (245.6 ± 3 mg/L) might be the main component that caused the observed 66 ± 2% decrease in anammox activity. The proliferation of denitrifying bacteria and sulfate-reducing bacteria induced by the organic compounds may have led to the observed decline in the abundance of the anammox bacterium Candidatus Kuenenia. The proposed strategy guaranteed the robust operation of the PN-anammox process and provides a promising approach for the sustainable treatment of incineration leachate.

Multidisciplinary characterization of nitrogen-removal granular sludge: A review of advances and technologies.

Nitrogen-removal granular sludge (NRGS) is a promising technology in wastewater treatment, with advantages of efficient nitrogen removal, less footprint, lower sludge production and energy consumption, and is a way for wastewater treatment plants to achieve carbon-neutrality. Aerobic granular sludge (AGS) and anammox granular sludge (AnGS) are two typical NRGS technologies that have attracted extensive attention. Mounting evidence has shown strong associations between NRGS properties and the status of NRGS systems; however, a holistic view is still missing. The aim of this article is to provide an overview of NRGS with an emphasis on characterization. Specifically, the integrated nitrogen transformation pathways inside NRGS and the performance of NRGS treating various wastewaters are discussed. NRGS properties are categorized as physical-, chemical-, biological- and systematical ones, presenting current advances and corresponding characterization technologies. Finally, the future prospects for furthering the mechanistic understanding and engineering application of NRGS are proposed. Overall, the technological advancements in characterization have greatly contributed to understanding NRGS properties, which are potential factors for optimizing the performance and evaluating the working status of NRGS. This review will provide guidance in characterizing NRGS properties and boost the introduction of novel characterization technologies.

Application of hydrophilic modified nylon fabric membrane in an anammox-membrane bioreactor: performance and fouling characteristics.

The membrane fouling is the main bottleneck hindering the wide applications of anammox-membrane bioreactor (MBR). In this study, surface-coating hydrophilic modification of the membrane using polyvinyl alcohol was applied in a granular anammox-MBR. Stable anammox performance of >77% total nitrogen removal efficiency was achieved in both original and modified MBRs, along with decreasing anammox granule size. The modified membrane exhibited superior flux performance, and the membrane foulants were reduced in the MBR operation. Specifically, the foulant formation rate (f) was 0.46 g·m-2·d-1 for the modified membrane with 100-µm coating thickness (M100) compared with 0.75 g·m-2·d-1 for the original membrane (M0). However, the fouling cycle of the modified membrane with 250-µm coating thickness (M250) was greatly shortened (5 days compared with 19 days for M0) and f increased to 1.25 g·m-2·d-1. Specially, the excess adhesion of exopolysaccharides and humic substances to the hydrophilic modified membrane changed the fouling layer structure and filtration resistance distribution, ultimately causing higher filtration resistance when coating thickness increased. Notably, the flux decline contribution of the concentration polarization was only 33.3% for M0, while it was 71.3% for M250. Finally, it was revealed that using a modified membrane increased the biological secretion rate of polysaccharide but decreased the protein bio-production rate, leading to a high PS (polysaccharide)/PN (protein) ratio in the MBR. The fouling mechanism of the hydrophilic modified membrane applied in anammox-MBR was proposed, and we highlight that the degree of hydrophilic modification is crucial to mitigating membrane fouling.

Function of Fe(III)-minerals in the enhancement of anammox performance exploiting integrated network and metagenomics analyses.

Iron is a recognized physiological requirement for microorganisms but, for anaerobic ammonium oxidation (anammox) bacteria, its role extends well beyond that of a nutritional necessity. In this study, the function of two typical Fe(III)-minerals (ferrihydrite and magnetite) in anammox processes was evaluated in the absence/presence of Fe(II) by integrated network and metagenomics analyses. Results showed that Fe-(III) minerals addition increased the activity of cellular processes and pathways associated with granule formation, enabling the peak values of particle size to increase by 144% and 115%, respectively. Notably, ferrihydrite (5 mM) enhanced nitrogen removal by 4.8% and 4.1%, respectively, in the short-term and long-term absence of Fe(II). Ferrihydrite also promoted the retention of anammox bacteria affiliated with phylum Planctomycetes in the reactor, contributing to an 11% higher abundance with ferrihydrite amendment when compared with the control (without iron additions) in the short-term absence of Fe(II). Network-based analyses revealed that ferrihydrite facilitated the microbial community to form densely clustered and complex topologies to improve resistance to environmental disturbance (i.e., Fe(II) deficiency), and effectively increased the underlying cooperation and facilitation in the community. Metagenomic analysis revealed that there was limited promotion of anammox central metabolism by the extra addition of Fe(III)-minerals in the presence of Fe(II), highlighting the poor utilization of Fe(III)-minerals by anammox bacteria under Fe(II) sufficiency. This study deepens our understanding of the function of Fe(III)-minerals in anammox systems at the community and functional level, and provides a fundamental basis for developing Fe-based anammox enhancement technologies.

COVID-19 inhibits spermatogenesis in the testes by inducing cellular senescence.

Introduction: COVID-19 (SARS-CoV-2) has been linked to organ damage in humans since its worldwide outbreak. It can also induce severe sperm damage, according to research conducted at numerous clinical institutions. However, the exact mechanism of damage is still unknown. Methods: In this study, testicular bulk-RNA-seq Data were downloaded from three COVID-19 patients and three uninfected controls from GEO to evaluate the effect of COVID-19 infection on spermatogenesis. Relative expression of each pathway and the correlation between genes or pathways were analyzed by bioinformatic methods. Results: By detecting the relative expression of each pathway and the correlation between genes or pathways, we found that COVID-19 could induce testicular cell senescence through MAPK signaling pathway. Cellular senescence was synergistic with MAPK pathway, which further affected the normal synthesis of cholesterol and androgen, inhibited the normal synthesis of lactate and pyruvate, and ultimately affected spermatogenesis. The medications targeting MAPK signaling pathway, especially MAPK1 and MAPK14, are expected to be effective therapeutic medications for reducing COVID-19 damage to spermatogenesis. Conclusion: These results give us a new understanding of how COVID-19 inhibits spermatogenesis and provide a possible solution to alleviate this damage.

MiR-629-5p Promotes Prostate Cancer Development and Metastasis by Targeting AKAP13.

Prostate cancer (PCa) has become the most frequently occurring cancer among western men according to the latest report, and patients' prognosis is often poor in the event of tumor progression, therefore, many researches are devoted to exploring the molecular mechanism of PCa metastasis. MicroRNAs (miRNA) have proved to play an important role in this process. In present study, by combining clinical samples with public databases, we found that miR-629-5p increased to varying degrees in primary localized PCa tissues and metastatic PCa tissues compared with adjacent normal tissues, and bioinformatics analysis suggested that high level of miR-629-5p was related to poor prognosis. Functionally, miR-629-5p drove PCa cell proliferation, migration and invasion in vitro, and promoted growth of PCa cells in vivo. Moreover, A-kinase Anchor Protein 13 (AKAP13) was screened as a direct target of miR-629-5p, that expression was negatively correlated with the malignant phenotype of tumor cells. In the end, through verification in clinical specimens, we found that AKAP13 could be independently used as a clinical prognostic indicator. Overall, the present study indicates that miR-629-5p plays an oncogenic role in PCa by targeting AKAP13, which provides a new idea for clinical diagnosis and treatment of complex refractory PCa.

The Correlation between PSCA Expression and Neuroendocrine Differentiation in Prostate Cancer.

The prostate stem cell antigen (PSCA), as a predominantly prostate-specific marker, is overexpressed in most prostate cancer specimens, is positively correlated with prostate cancer androgen independence, and has the potential to be treated with castration-resistant prostate cancer (CRPC) as a gene therapy target. Using the typical androgen deprivation therapy, most tumors will progress to CRPC, as well as develop into neuroendocrine prostate cancer (NEPC) characterized by the expression of neuroendocrine markers such as enolase 2 (NSE). Our study was aimed at investigating the expressions of PSCA and NSE and the relationship between the two markers, as well as the correlation between the PSCA and NSE expressions and the clinicopathological parameters in prostate cancer specimens from 118 patients by using immunohistochemistry. Our results demonstrated that the PSCA and NSE protein expressions did not correlate with the prostate cancer patients' age or the hormone therapy but showed a significant correlation with the pathological tumor stage of prostate cancer, the Gleason score, and the presence of metastasis. There is a positive association between PSCA and NSE but a negative one between the prostate-specific antigen (PSA) and PSCA or between PSA and NSE. High PSCA and NSE expressions correlated with a poor prognosis in prostate cancer patients. PSCA may play an important role in the progression of neuroendocrine prostate cancer (NEPC).

Organic compounds evolution and sludge properties variation along partial nitritation and subsequent anammox processes treating reject water.

Reject water contains complex components of organic compounds, which have significant influences on the nitrogen removal performance when treated using biological autotrophic nitrogen removal technology. In this study, a two-stage partial nitritation (PN)-anammox (floc-granule) system was established to treat reject water (COD/NH4+-N = 0.97 ± 0.15), and the evolution of organic compounds along PN and annamox bioreactors was investigated using gas chromatography-mass spectrometry and excitation-emission matrix. Also, the variation of PN and anammox sludge properties relating to COD reduction was examined. The PN-anammox system removed approximately 80% of total inorganic nitrogen and COD with hydraulic reaction time of 16 h. The influent organics (330-600 mg COD/L) in reject water were primarily composed of volatile, protein-like and humic acid-like organic compounds. PN process contributed 53 ± 18% of the overall COD removal, primarily including oxygen-containing organics (e.g. phenol), proteins and humic acids. Anammox process contributed 22 ± 15% of the overall COD removal, but large molecule acids (e.g. lactic acid) and small molecule alcohols (e.g. glycerol) were reoccurred, contributing to the effluent COD with recalcitrant hydrocarbons (e.g. n-Octadecane). Reject water increased the extracellular proteins/polysaccharides ratio of PN and anammox sludge, promoting the adsorption and degradation of organic compounds. High-throughput sequencing results showed that denitrifying bacteria of Ottowia increased from 0.03% to 14.4% in PN reactor, and of Denitratisoma increased from 9.6% to 15.4% in anammox reactor. The occurrence of these denitrifiers might mitigate the negative impact of organics to functional organisms. This study highlights the organics fate during PN-anammox treatment system, which is important to maintain the robust nitrogen removal when treating organics-containing and high ammonium concentration wastewater.

The m6A methylation regulator-based signature for predicting the prognosis of prostate cancer.

Aim: To construct a survival prediction signature for prostate cancer (PC) based on the RNA N6-methyladenosine (m6A) methylation regulator. Materials & methods: This paper explores the interaction network of differentially expressed m6A RNA methylation regulators in PC by Pearson correlation analysis. Univariate Cox risk regression and LASSO regression analysis were used to construct a predictive signature of PC. Kaplan-Meier survival analysis compared the overall survival of the high- and low-risk groups. Results & Conclusion: We first constructed a prognostic two gene signature for PC based on the m6A RNA methylation regulators MRTTL14 and YTHDF2. The interaction network of m6A RNA methylation regulators in PC was also established.

Chopper-modulated gas chromatography electroantennography enabled using high-temperature MEMS flow control device.

We report the design, fabrication and characterization of a microelectromechanical systems (MEMS) flow control device for gas chromatography (GC) with the capability of sustaining high-temperature environments. We further demonstrate the use of this new device in a novel MEMS chopper-modulated gas chromatography-electroantennography (MEMS-GC-EAG) system to identify specific volatile organic compounds (VOCs) at extremely low concentrations. The device integrates four pneumatically actuated microvalves constructed via thermocompression bonding of the polyimide membrane between two glass substrates with microstructures. The overall size of the device is 32 mm×32 mm, and it is packaged in a 50 mm×50 mm aluminum housing that provides access to the fluidic connections and allows thermal control. The characterization reveals that each microvalve in the flow control chip provides an ON to OFF ratio as high as 1000:1. The device can operate reliably for more than 1 million switching cycles at a working temperature of 300 °C. Using the MEMS-GC-EAG system, we demonstrate the successful detection of cis-11-hexadecenal with a concentration as low as 1 pg at a demodulation frequency of 2 Hz by using an antenna harvested from the male Helicoverpa Virescens moth. In addition, 1 µg of a green leafy volatile (GLV) is barely detected using the conventional GC-EAG, while MEMS-GC-EAG can readily detect the same amount of GLV, with an improvement in the signal-to-noise ratio (SNR) of ~22 times. We expect that the flow control device presented in this report will allow researchers to explore new applications and make new discoveries in entomology and other fields that require high-temperature flow control at the microscale.

G-Fresnel smartphone spectrometer.

We report a smartphone spectrometer with nanometer resolution working in the visible range. A G-Fresnel device with the dual functionality of focusing and dispersion is used to enable miniaturization. Proof of principle application to Bradford assay of protein concentration is also demonstrated.

On-demand one-step synthesis of monodisperse functional polymeric microspheres with droplet microfluidics.

A simple and robust method for one-step synthesis of monodisperse functional polymeric microspheres was established by generation of reversed microemulsion droplets in aqueous phase inside microfluidic chips and controlled evaporation of the organic solvent. Using this method, water-soluble nanomaterials can be easily encapsulated into biodegradable Poly(D,L-lactic-co-glycolic acid) (PLGA) to form functional microspheres. By controlling the flow rate of microemulsion phase, PLGA polymeric microspheres with narrow size distribution and diameters in the range of ∼50-100 µm were obtained. As a demonstration of the versatility of the approach, high-quality fluorescent CdTe:Zn(2+) quantum dots (QDs) of various emission spectra, superparamagnetic Fe3O4 nanoparticles, and water-soluble carbon nanotubes (CNTs) were used to synthesize fluorescent PLGA@QDs, magnetic PLGA@Fe3O4, and PLGA@CNTs polymeric microspheres, respectively. In order to show specific applications, the PLGA@Fe3O4 were modified with polydopamine (PDA), and then the silver nanoparticles grew on the surfaces of the PLGA@Fe3O4@PDA polymeric microspheres by reducting the Ag(+) to Ag(0). The as-prepared PLGA@Fe3O4@PDA-Ag microspheres showed a highly efficient catalytic reduction of the 4-nitrophenol, a highly toxic substance. The monodisperse uniform functional PLGA polymeric microspheres can potentially be critically important for multiple biomedical applications.

Separation and purification of both tea seed polysaccharide and saponin from camellia cake extract using macroporous resin.

A novel method to separate and purify tea seed polysaccharide and tea seed saponin from camellia cake extract by macroporous resin was developed. Among four kinds of resins (AB-8, NKA-9, XDA-6, and D4020) tested, AB-8 macroporous resin possessed optimal separating capacity for the two substances and thus was selected for the separation, in which deionized water was used to elute tea seed polysaccharide, 0.25% NaOH solution to remove the undesired pigments, and 90% ethanol to elute tea seed saponin. Further dynamic adsorption/desorption experiments on AB-8 resin-based column chromatography were conducted to obtain the optimal parameters. Under optimal dynamic adsorption and desorption conditions, 18.7 and 11.8% yield of tea seed polysaccharide and tea seed saponin were obtained with purities of 89.2 and 96.0%, respectively. The developed method provides a potential approach for the large-scale production of tea seed polysaccharide and tea seed saponin from camellia cake.

Separable bilayer microfiltration device for viable label-free enrichment of circulating tumour cells.

The analysis of circulating tumour cells (CTCs) in cancer patients could provide important information for therapeutic management. Enrichment of viable CTCs could permit performance of functional analyses on CTCs to broaden understanding of metastatic disease. However, this has not been widely accomplished. Addressing this challenge, we present a separable bilayer (SB) microfilter for viable size-based CTC capture. Unlike other single-layer CTC microfilters, the precise gap between the two layers and the architecture of pore alignment result in drastic reduction in mechanical stress on CTCs, capturing them viably. Using multiple cancer cell lines spiked in healthy donor blood, the SB microfilter demonstrated high capture efficiency (78-83%), high retention of cell viability (71-74%), high tumour cell enrichment against leukocytes (1.7-2 × 10(3)), and widespread ability to establish cultures post-capture (100% of cell lines tested). In a metastatic mouse model, SB microfilters successfully enriched viable mouse CTCs from 0.4-0.6 mL whole mouse blood samples and established in vitro cultures for further genetic and functional analysis. Our preliminary studies reflect the efficacy of the SB microfilter device to efficiently and reliably enrich viable CTCs in animal model studies, constituting an exciting technology for new insights in cancer research.