Optimization of microbial fuel cell performance application to high sulfide industrial wastewater treatment by modulating microbial function.

Microbial fuel cells (MFCs) are innovative eco-friendly technologies that advance a circular economy by enabling the conversion of both organic and inorganic substances in wastewater to electricity. While conceptually promising, there are lingering questions regarding the performance and stability of MFCs in real industrial settings. To address this research gap, we investigated the influence of specific operational settings, regarding the hydraulic retention time (HRT) and organic loading rate (OLR) on the performance of MFCs used for treating sulfide-rich wastewater from a canned pineapple factory. Experiments were performed at varying hydraulic retention times (2 days and 4 days) during both low and high seasonal production. Through optimization, we achieved a current density generation of 47±15 mA/m2, a COD removal efficiency of 91±9%, and a sulfide removal efficiency of 86±10%. Microbiome analysis revealed improved MFC performance when there was a substantial presence of electrogenic bacteria, sulfide-oxidizing bacteria, and methanotrophs, alongside a reduced abundance of sulfate-reducing bacteria and methanogens. In conclusion, we recommend the following operational guidelines for applying MFCs in industrial wastewater treatment: (i) Careful selection of the microbial inoculum, as this step significantly influences the composition of the MFC microbial community and its overall performance. (ii) Initiating MFC operation with an appropriate OLR is essential. This helps in establishing an effective and adaptable microbial community within the MFCs, which can be beneficial when facing variations in OLR due to seasonal production changes. (iii) Identifying and maintaining MFC-supporting microbes, including those identified in this study, should be a priority. Keeping these microbes as an integral part of the system's microbial composition throughout the operation enhances and stabilizes MFC performance.

Detection of high-risk HPV 16 genotypes in cervical cancers using isothermal DNA amplification with electrochemical genosensor.

Cervical cancer emerges as the third most prevalent types of malignancy among women on a global scale. Cervical cancer is significantly associated with the persistent infection of human papillomavirus (HPV) type 16. The process of diagnosing is crucial in order to prevent the progression of a condition into a malignant state. The early detection of cervical cancer through initial stage screening is of the utmost significance in both the prevention and effective management of this disease. The present detection methodology is dependent on quantitative polymerase chain reaction (qPCR), which necessitates the use of a costly heat cycler instrument. In this study, we report the development of an electrochemical DNA biosensor integrated with an isothermal recombinase polymerase amplification (RPA) reaction for the detection and identification of the high-risk HPV-16 genotype. The electrochemical biosensor exhibited a high degree of specificity and sensitivity, as evidenced by its limit of detection (LOD) of 0.23 copies/µL of HPV-16 DNA. The validity of this electrochemical platform was confirmed through the analysis of 40 cervical tissues samples, and the findings were consistent with those obtained through polymerase chain reaction (PCR) testing. Our straightforward electrochemical detection technology and quick turnaround time at 75 min make the assay suitable for point-of-care testing in low-resource settings.

Effect of propionate-cultured sludge augmentation on methane production from upflow anaerobic sludge blanket systems treating fresh landfill leachate.

This research investigates the effect of propionate-cultured sludge augmentation on methane (CH4) production from upflow anaerobic sludge blanket systems (UASB) treating fresh landfill leachate. In the study, both UASB reactors (UASB 1 and UASB 2) contained acclimatized seed sludge, and UASB 2 was augmented with propionate-cultured sludge. The organic loading rate (OLR) was varied between 120.6, 84.4, 48.2, and 12.0 gCOD/L·d. The experimental results indicated that the optimal OLR of UASB 1 (non-augmentation) was 48.2 gCOD/L·d, achieving the CH4 production of 4019 mL/d. Meanwhile, the optimal OLR of UASB 2 was 12.0 gCOD/L·d, achieving the CH4 yield of 6299 mL/d. The dominant bacterial community in the propionate-cultured sludge included the genera Methanothrix, Methanosaeta, Methanoculleus, Syntrophobacter, Smithella, Pelotomamulum, which are the VFA-degrading bacteria and methanogens responsible for unblocking the CH4 pathway bottleneck. Essentially, the novelty of this research lies in the use of propionate-cultured sludge to augment the UASB reactor in order to enhance CH4 production from fresh landfill leachate.

Propionate-cultured sludge bioaugmentation to enhance methane production and micropollutant degradation in landfill leachate treatment.

This research investigates the use of propionate-cultured sludge to enhance methane (CH4) production and micropollutant biodegradation in biochemical methane potential (BMP) experiment treating landfill leachate. The experiments were carried out using non-acclimatized and acclimatized seed sludge with variable food to microorganism ratios of 1:1 and 1:2. Under the propionate-cultured sludge bioaugmentation, the concentrations of propionate-cultured sludge were varied between 10, 20, and 30 % (v/v). The acclimatized seed sludge exhibited high microbial abundance and diversity which promoted the CH4 production and micropollutant biodegradation. The modified Gompertz model indicated that the optimal condition was the acclimatized seed sludge with 30% (v/v) propionate-cultured sludge, achieving the lag time (λ), maximum CH4 production rate (Rmax), and maximum CH4 potential yield (Pmax) of 0.57 day, 17.35 NmL/h, and 140.58 NmL/g COD. The research novelty lies in the use of propionate-cultured sludge bioaugmentation in landfill leachate treatment to enhance CH4 production and micropollutant biodegradation.

Treatment efficiency and greenhouse gas emissions of non-floating and floating bed activated sludge system with acclimatized sludge treating landfill leachate.

This research investigates the treatment efficiency and greenhouse gas (GHG) emissions of non-floating and floating bed AS systems with acclimatized sludge treating landfill leachate. The GHGs under study included carbon dioxide (CO2), methane (CH4), and nitrous oxide (N2O). The non-floating and floating bed AS systems were operated in parallel with identical landfill leachate influent under different hydraulic retention time (HRT) conditions (24, 18, and 12 h). The experimental results showed that the treatment efficiency of organic compounds under 24 h HRT of both systems (90 - 98%) were insignificantly different, while the nutrient removal efficiency of both systems were between 54 and 98 %. The treatment efficiency of the floating bed AS system, despite shorter HRT, remained relatively unchanged due to an abundance of effective bacteria residing in the floating media. The CO2 emissions were insignificantly different between both AS systems under all HRT conditions (22 - 26.3 µmol/cm2.min). The CO2 emissions were positively correlated with organic loading but inversely correlated with HRT. The CH4 emissions were positively correlated with HRT (26.3 µmol/cm2.min under 24 h HRT of the floating bed AS system). The N2O emissions were positively correlated with nitrogen loading, and the N2O emissions from the floating bed AS system were lower due to an abundance of N2O-reducing bacteria. The floating media enhanced the biological treatment efficiency while maintaining the bacterial community in the system. However, the floating media promoted CH4 production under anoxic conditions. The originality of this research lies in the use of floating media in the biological treatment system to mitigate GHG emissions, unlike existing research which focused primarily on enhancement of the treatment efficiency.

Rapid electrochemical detection of coronavirus SARS-CoV-2.

Coronavirus disease 2019 (COVID-19) is a highly contagious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Diagnosis of COVID-19 depends on quantitative reverse transcription PCR (qRT-PCR), which is time-consuming and requires expensive instrumentation. Here, we report an ultrasensitive electrochemical biosensor based on isothermal rolling circle amplification (RCA) for rapid detection of SARS-CoV-2. The assay involves the hybridization of the RCA amplicons with probes that were functionalized with redox active labels that are detectable by an electrochemical biosensor. The one-step sandwich hybridization assay could detect as low as 1 copy/µL of N and S genes, in less than 2 h. Sensor evaluation with 106 clinical samples, including 41 SARS-CoV-2 positive and 9 samples positive for other respiratory viruses, gave a 100% concordance result with qRT-PCR, with complete correlation between the biosensor current signals and quantitation cycle (Cq) values. In summary, this biosensor could be used as an on-site, real-time diagnostic test for COVID-19.

Diverse Microbial Community Profiles of Propionate-Degrading Cultures Derived from Different Sludge Sources of Anaerobic Wastewater Treatment Plants.

Anaerobic digestion (AD) has been used for wastewater treatment and production of renewable energy or biogas. Propionate accumulation is one of the important problems leading to an unstable system and low methane production. Revealing propionate-degrading microbiome is necessary to gain a better knowledge for alleviation of the problem. Herein, we systematically investigated the propionate-degrading cultures enriched from various anaerobic sludge sources of agro-industrial wastewater treatment plants using 16S rRNA gene sequencing. Different microbial profiles were shown even though the methanogenic activities of all cultures were similar. Interestingly, non-classical propionate-degrading key players Smithella, Syntrophomonas, and Methanosaeta were observed as common prevalent taxa in our enriched cultures. Moreover, different hydrogenotrophic methanogens were found specifically to the different sludge sources. The enriched culture of high salinity sludge showed a distinct microbial profile compared to the others, containing mainly Thermovirga, Anaerolinaceae, Methanosaeta, Syntrophobactor, and Methanospirillum. Our microbiome analysis revealed different propionate-degrading community profiles via mainly the Smithella pathway and offers inside information for microbiome manipulation in AD systems to increase biogas production corresponding to their specific microbial communities.

Effect of hydraulic retention time on micropollutant biodegradation in activated sludge system augmented with acclimatized sludge treating low-micropollutants wastewater.

This research investigates the effect of hydraulic retention time (HRT) on micropollutant biodegradation of two-stage activated sludge (AS) system augmented with acclimatized sludge treating low-micropollutants wastewater. The experimental wastewater was a mixture of landfill leachate and agriculture wastewater, and HRT was varied between 24, 18, and 12 h. The results showed that, under 24 h HRT, the micropollutant biodegradation efficiencies were 87-93% for bisphenol A (BPA), 2,6-di-tert-butyl-phenol (2,6-DTBP), di-butyl-phthalate (DBP), di-(ethylhexyl)-phthalate (DEHP); 75-81% for carbamazepine (CBZ), diclofenac (DCF); and 88% for N,N-diethylmeta-toluamide (DEET). The degradation efficiencies were similar under 18 h HRT: 87-93% for BPA, 2,6-DTBP, DBP, DEHP; 75-80% for CBZ, DCF; and 80% for DEET. However, the efficiencies substantially declined under 12 h HRT: 71-93%, 55-60%, and 50%, respectively. Importantly, the findings revealed that HRT plays a crucial part in micropollutant biodegradation of bioaugmented AS system. More specifically, too short an HRT (12 h) results in low micropollutant removal efficiency, and too long an HRT (24 h) contributes to low daily throughput and high treatment operation cost. As a result, moderate HRT (18 h) is operationally and economically optimal for bioaugmented AS system treating low-micropollutants wastewater.

Potentially toxic elements to maize in agricultural soils-microbial approach of rhizospheric and bulk soils and phytoaccumulation.

Maize fields near Mae Tao Creek in Pha Te Village, Tak Province, Thailand are contaminated with Zn, Cd, and Pb. This research studied the interaction between levels of the metals contaminating the soil and maize development, heavy metal accumulation in the seeds, and the soil bacterial community structure. Our field experiment was carried out in five plots with metal contents that gradually decreased from a high level near the creek to a lower level further into the land: Zn 380-4883 mg kg-1, Cd 6-85 mg kg-1, and Pb 34-154 mg kg-1. Cultivation and isolation on nutrient agar (NA) was utilized to study the culturable bacterial community, and polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) was utilized for the unculturable bacterial communities. All statistical analyses clearly indicated that rainfall and irrigation were the main factors affecting total Zn concentration and bioavailable Zn, Cd, and Pb in the field. The variation in the contents of the heavy metals was weakly correlated with the culturable bacterial community indices (Shannon-Wiener, evenness and richness), but the contents resulted in a difference in the overall diversity of the bacteria in the soil. The richness, numbers of culturable rhizobacteria, and maize growth stage significantly affected the amount of Zn and Cd that accumulated in the roots. In addition, maize accumulated a high level of Zn in the seeds, while the low contents of Cd and Pb in the seeds were below our limit of detection. The results obtained could be informative for the management of maize cultivation in the area.

Use of aged sludge bioaugmentation in two-stage activated sludge system to enhance the biodegradation of toxic organic compounds in high strength wastewater.

This research investigates the toxic organic compounds biodegradation efficiency of two-stage activated sludge systems with (bioaugmented) and without aged sludge bioaugmentation (non-bioaugmented). The influent was a mixture of leachate and agriculture wastewater (1:1, v/v), used as the representative high strength wastewater. The bioaugmented and non-bioaugmented systems were operated in parallel, with three levels (low, moderate, and high) of concentrations of organics, nitrogen, and toxic organic compounds in the influent (conditions 1, 2, and 3). The results showed that both systems could efficiently degrade the organic compounds. Nevertheless, the toxic organic compounds biodegradation efficiency of the bioaugmented system was higher than that of the non-bioaugmented one. The bioaugmentation enhanced the overall removal efficiency under conditions 1 and 2. However, the bioaugmented system became less effective under condition 3. Further analysis indicated that the bacterial groups essential to the toxic organic compounds biodegradation were abundant in the aged sludge, including heterotrophic bacteria, heterotrophic nitrifying bacteria, and nitrifying bacteria. The abundance of the effective bacteria improved the biodegradation and wastewater treatment performance of the bioaugmented system. In essence, the aged sludge bioaugmentation is a viable and eco-friendly solution to improving the treatment efficiency of the biological activated sludge system, despite limited biodegradation efficiency in an elevated compounds-concentration environment.

Toxic compounds biodegradation and toxicity of high strength wastewater treated under elevated nitrogen concentration in the activated sludge and membrane bioreactor systems.

This research has assessed the removal efficiencies of toxic compounds in the high strength wastewater (the leachate and agriculture wastewater mixture) using the activated sludge (AS) and membrane bioreactor (MBR) technologies under two carbon to nitrogen (C/N) ratios (C/N 14 and 6) and two toxic compounds concentrations (8-396µg/L and 1000µg/L). In addition, the toxicity evaluations of the AS and MBR effluents to the aquatic environment were undertaken at five effluent dilution ratios (10, 20, 30, 50 and 70% v/v). The findings indicate that the AS treatment performance could be enhanced by the elevation of the nitrogen concentration. Specifically, the C/N 6 environment helps promote the bacterial growth, particularly heterotrophic nitrifying bacteria (HNB) and nitrifying bacteria (NB), which produce the enzymes crucial to the toxic compounds degradation. The improved biodegradation makes the effluents less toxic to the aquatic environment, as evidenced by the lower mortality rates of both experimental fish species raised in the nitrogen-elevated diluted AS effluents. On the other hand, the elevated nitrogen concentration minimally enhances the MBR treatment performance, given the fact that the MBR technology is in itself a biological treatment scheme with very high compounds removal capability. Despite its lower toxic compounds removal efficiency, the AS technology is simple, inexpensive and operationally-friendly, rendering the system more applicable to the treatment operation constrained by the financial, manpower and technological considerations.

Diversity of purple nonsulfur bacteria in shrimp ponds with varying mercury levels.

This research aimed to study the diversity of purple nonsulfur bacteria (PNSB) and to investigate the effect of Hg concentrations in shrimp ponds on PNSB diversity. Amplification of the pufM gene was detected in 13 and 10 samples of water and sediment collected from 16 shrimp ponds in Southern Thailand. In addition to PNSB, other anoxygenic phototrophic bacteria (APB) were also observed; purple sulfur bacteria (PSB) and aerobic anoxygenic phototrophic bacteria (AAPB) although most of them could not be identified. Among identified groups; AAPB, PSB and PNSB in the samples of water and sediment were 25.71, 11.43 and 8.57%; and 27.78, 11.11 and 22.22%, respectively. In both sample types, Roseobacter denitrificans (AAPB) was the most dominant species followed by Halorhodospira halophila (PSB). In addition two genera, observed most frequently in the sediment samples were a group of PNSB (Rhodovulum kholense, Rhodospirillum centenum and Rhodobium marinum). The UPGMA dendrograms showed 7 and 6 clustered groups in the water and sediment samples, respectively. There was no relationship between the clustered groups and the total Hg (HgT) concentrations in the water and sediment samples used (<0.002-0.03 µg/L and 35.40-391.60 µg/kg dry weight) for studying the biodiversity. It can be concluded that there was no effect of the various Hg levels on the diversity of detected APB species; particularly the PNSB in the shrimp ponds.

ANAMMOX-like performances for nitrogen removal from ammonium-sulfate-rich wastewater in an anaerobic sequencing batch reactor.

Ammonium removal by the ANaerobic AMonium OXidation (ANAMMOX) process was observed through the Sulfate-Reducing Ammonium Oxidation (SRAO) process. The same concentration of ammonium (100 mg N L(-1)) was applied to two anaerobic sequencing batch reactors (AnSBRs) that were inoculated with the same activated sludge from the Vermicelli wastewater treatment process, while nitrite was fed in ANAMMOX and sulfate in SRAO reactors. In SRAO-AnSBR, in substrates that were fed with a ratio of NH4(+)/SO4(2-) at 1:0.4 ± 0.03, a hydraulic retention time (HRT) of 48 h and without sludge draining, the Ammonium Removal Rate (ARR) was 0.02 ± 0.01 kg N m(-3).d(-1). Adding specific ANAMMOX substrates to SRAO-AnSBR sludge in batch tests results in specific ammonium and nitrite removal rates of 0.198 and 0.139 g N g(-1) VSS.d, respectively, indicating that the ANAMMOX activity contributes to the removal of ammonium in the SRAO process using the nitrite that is produced from SRAO. Nevertheless, the inability of ANAMMOX to utilize sulfate to oxidize ammonium was also investigated in batch tests by augmenting enriched ANAMMOX culture in SRAO-AnSBR sludge and without nitrite supply. The time course of sulfate in a 24-hour cycle of SRAO-AnSBR showed an increase in sulfate after 6 h. For enriched SRAO culture, the uptake molar ratio of NH4(+)/SO4(2-) at 8 hours in a batch test was 1:0.82 lower than the value of 1:0.20 ± 0.09 as obtained in an SRAO-AnSBR effluent, while the stoichiometric ratio of 1:0.5 that includes the ANAMMOX reaction was in this range. After a longer operation of more than 2 years without sludge draining, the accumulation of sulfate and the reduction of ammonium removal were observed, probably due to the gradual increase in the sulfur denitrification rate and the competitive use of nitrite with ANAMMOX. The 16S rRNA gene PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) and PCR cloning analyses resulted in the detection of the ANAMMOX bacterium (Candidatus Brocadia sinica JPN1) Desulfacinum subterraneum belonging to the genus Desulfacinum and bacteria that are involved in sulfur metabolism (Pseudomonas aeruginosa strain SBTPe-001 and Paracoccus denitrificans strain IAM12479) in SRAO-AnSBR.

Microbial communities and their performances in anaerobic hybrid sludge bed-fixed film reactor for treatment of palm oil mill effluent under various organic pollutant concentrations.

The anaerobic hybrid reactor consisting of sludge and packed zones was operated with organic pollutant loading rates from 6.2 to 8.2 g COD/L day, composed mainly of suspended solids (SS) and oil and grease (O&G) concentrations between 5.2 to 10.2 and 0.9 to 1.9 g/L, respectively. The overall process performance in terms of chemical oxygen demands (COD), SS, and O&G removals was 73, 63, and 56%, respectively. When the organic pollutant concentrations were increased, the resultant methane potentials were higher, and the methane yield increased to 0.30 L CH4/g COD(removed). It was observed these effects on the microbial population and activity in the sludge and packed zones. The eubacterial population and activity in the sludge zone increased to 6.4 × 109 copies rDNA/g VSS and 1.65 g COD/g VSS day, respectively, whereas those in the packed zone were lower. The predominant hydrolytic and fermentative bacteria were Pseudomonas, Clostridium, and Bacteroidetes. In addition, the archaeal population and activity in the packed zone were increased from to 9.1 × 107 copies rDNA/g VSS and 0.34 g COD-CH4/g VSS day, respectively, whereas those in the sludge zone were not much changed. The most represented species of methanogens were the acetoclastic Methanosaeta, the hydrogenotrophic Methanobacterium sp., and the hydrogenotrophic Methanomicrobiaceae.

Molecular monitoring of microbial population dynamics during operational periods of anaerobic hybrid reactor treating cassava starch wastewater.

This study characterized the microbial community and population dynamics in an anaerobic hybrid reactor (AHR) treating cassava starch wastewater. Methanogens and nonmethanogens were followed during the start-up and operation of the reactor, and linked to operational and performance data. Biomass samples taken from the sludge bed and packed bed zones of the AHR at intervals throughout the operational period were examined by 16S rRNA fluorescence in situ hybridization (FISH). The start-up seed and the reactor biomass were sampled during the feeding of the wastewater with a chemical oxygen demand (COD) value of 8 g L(-1) and a hydraulic retention time (HRT) of 8 days. These samples were characterized by the predominance of cells with long-rod morphology similar to Methanosaeta spp. Following a sharp operational change, accomplished by increasing the COD concentration of the organic influent from 8 to 10 g L(-1) and reducing the HRT from 8 to 5 days, there was a doubling of the organic loading rate, a reduction of the COD removal efficiency, as well as decreased methane content in the biogas and an accumulation of total volatile acids in the reactor. Moreover, this operational change resulted in a significant population shift from long-rod Methanosaeta-like cells to tetrad-forming Methanosarcina-like cells. The distributions of microbial populations involved in different zones of the AHR were determined. The results showed that nonmethanogens became the predominant population in both sludge and the packed bed zone. However, the percentage of methanogens in the packed bed zone was higher than that in the sludge bed zone. This higher percentage of methanogens was likely caused by the fact that the packed bed zone provided a suitable environmental condition with an appropriate nutrient availability for methanogen growth.

The serine protease inhibitor trappin-2 is present in cartilage and synovial fluid in osteoarthritis.

OBJECTIVE: Trappins are small serine protease inhibitors bound to extracellular matrix (ECM) through the actions of transglutaminase (TGase) enzymes. Trappin-2 is present in many tissues and is upregulated at sites of injury. In osteoarthritis (OA), serine proteases contribute to articular cartilage destruction, and TGase activity is increased. Yet little is known about matrix-bound serine protease inhibitors or TGase substrates in articular cartilage. Our purpose was to determine if trappin-2 was present in OA cartilage and synovial fluid (SF). METHODS: OA knee articular cartilage and SF were assayed for trappin-2 protein by Western blotting, ELISA, and immunohistochemistry. Trappin-2 mRNA was detected with RT-PCR. The ECM components bound to trappin-2 were identified by 2-D gel electrophoresis and peptide fingerprinting. RESULTS: Trappin-2 was detectable in OA articular cartilage extracts, cultured chondrocytes, conditioned media, and SF by Western blotting. OA cartilage protein extracts contained significantly higher quantities of trappin-2 than normal cartilage protein extracts (22.98 +/- 1.28 ng/mg wet weight vs 14.97 +/- 1.92 ng/mg wet weight; p < 0.01). RT-PCR confirmed the presence of trappin-2 mRNA in OA chondrocytes. Immunohistochemical studies of OA cartilage revealed trappin-2 protein in chondrocytes. Peptide mapping of trappin-2 binding partners showed that fibromodulin was bound to trappin-2 in cartilage. CONCLUSION: We confirmed the presence of trappin-2 in OA cartilage and SF. Elevated levels of TGase activity in OA cartilage may increase levels of this serine protease inhibitor in response to injury.

Use of an alternative Archaea-specific probe for methanogen detection.

An alternative 16S rRNA-targeted oligonucleotide probe specific for Archaea was developed and used for detection of methanogens in anaerobic reactors. The designed probe was checked for its specificity by computer-aided comparative sequence analysis. For in situ application, optimal stringency conditions were adjusted by performing whole cell hybridization using target and nontarget organisms. Anaerobic sludge samples were examined by in situ hybridization for methanogenic populations. The relative abundance of methanogens was monitored with epifluorescence microscopy. Individual cells could be visualized with strong fluorescence signals after hybridization with the newly developed probe.