scholarly journals Applicability of infectivity assay for the quantification of infectious human adenovirus genotype 5 in UV-irradiated wastewater

2020 ◽  
Author(s):  
Surapong Rattanakul ◽  
Kumiko Oguma
Keyword(s):  
A549 Cells ◽  
Domestic Wastewater ◽  
Human Adenovirus ◽  
Host Cells ◽  
Uv Disinfection ◽  
Oh Radicals ◽  
Infectivity Assay ◽  
Water Matrix ◽  
Wastewater Samples ◽  
Inactivation Efficiency

Abstract The use of infectivity assays in domestic wastewater samples are limited because of the concerns on cytotoxicity to host cells, thus, the UV inactivation efficiency of human adenoviruses (HAdVs) in wastewater remains unclear. In this study, a human adenovirus genotype five (HAdV-5) host cell line (A549 cells) was incubated with wastewater at varied dilutions from 1:1.5 to 1:4 (the ratio of wastewater to a mixture of wastewater and the cell culture medium) and the cytotoxicity was assessed by the cell morphology and viability. No change was observed in either cell viability or morphology in comparison with control samples, even at lowest dilution of 1:1.5, indicating the dilution allowed infectivity assays. The minimal degree of dilution to avoid cytotoxicity may differ with different water matrix. Consequently, the technique was applied to quantify spiked HAdV-5 after the UV disinfection of wastewater. A significant increase in UV disinfection efficiency was noted in wastewater and hydroxyl radicals (OH•) produced by the photosensitization of dissolved organic matter were suggested to be responsible for the enhancement. This study indicated that dilution can be a simple solution to avoid cytotoxicity, and UV inactivation may be enhanced in wastewater due to OH• radicals produced by UV radiation.

Behaviour of Coli Phages in Oxidation Ponds

1985 ◽  
Vol 17 (10) ◽  
pp. 219-227 ◽  
Author(s):  
T. Omura ◽  
H. K. Shin ◽  
A. Ketratanakul
Keyword(s):  
Laboratory Experiments ◽  
Field Investigation ◽  
Host Cells ◽  
Coliform Bacteria ◽  
Alkaline Ph ◽  
Viral Inactivation ◽  
Effects Of Temperature ◽  
Oxidation Ponds ◽  
Incubation Temperatures ◽  
Inactivation Efficiency

Coliphages are among the most promising indicators of viral inactivation efficiency of wastewater treatment. Therefore, it is important to investigate the behaviour of coliphages in oxidation ponds from the viewpoint of predicting the inactivation of infectious viruses. In this study, numbers of coliphages were measured in oxidation ponds consisting of a series of facultative and maturation ponds. In parallel with this investigation, the effects of temperature and pH on the behaviour of coliphages were examined in the laboratory, employing three species of coliform bacteria as host cells. The field investigation showed that there was positive correlation between counts of coliphages and those of coliform bacteria, and that more than 99% of coliphages were inactivated. The inactivation efficiency of coliphages in the facultative pond was much higher than in the maturation pond. The results of the laboratory experiments indicated that at 30°C more than 99% of the coliform group were destroyed in 7 days of incubation and that coliphages counts increased from 105/100 ml to 107/100 ml with a lag time of 3 days. Greater reduction of the coliform count was obtained at higher incubation temperatures. It was observed that the coliphages possessed greater ability to attack coliform bacteria at acidic rather than alkaline pH.

scholarly journals Woven-Fiber Microfiltration (WFMF) and Ultraviolet Light Emitting Diodes (UV LEDs) for Treating Wastewater and Septic Tank Effluent

Water ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 1564
Author(s):  
Sara Beck ◽  
Poonyanooch Suwan ◽  
Thusitha Rathnayeke ◽  
Thi Nguyen ◽  
Victor Huanambal-Sovero ◽  
...  
Keyword(s):  
Wastewater Treatment ◽  
Light Emitting Diodes ◽  
Domestic Wastewater ◽  
Septic Tank ◽  
Uv Disinfection ◽  
Small Community ◽  
Photochemical Process ◽  
Light Emitting ◽  
Decentralized Wastewater Treatment ◽  
Septic Tank Effluent

Decentralized wastewater treatment systems enable wastewater to be treated at the source for cleaner discharge into the environment, protecting public health while allowing for reuse for agricultural and other purposes. This study, conducted in Thailand, investigated a decentralized wastewater treatment system incorporating a physical and photochemical process. Domestic wastewater from a university campus and conventional septic tank effluent from a small community were filtered through a woven-fiber microfiltration (WFMF) membrane as pretreatment for ultraviolet (UV) disinfection. In domestic wastewater, WFMF reduced TSS (by 79.8%), turbidity (76.5%), COD (38.5%), and NO3 (41.4%), meeting Thailand irrigation standards for every parameter except BOD. In septic tank effluent, it did not meet Thailand irrigation standards, but reduced TSS (by 77.9%), COD (37.6%), and TKN (13.5%). Bacteria (total coliform and Escherichia coli) and viruses (MS2 bacteriophage) passing through the membrane were disinfected by flow-through UV reactors containing either a low-pressure mercury lamp or light-emitting diodes (LEDs) emitting an average peak wavelength of 276 nm. Despite challenging and variable water quality conditions (2% < UVT < 88%), disinfection was predictable across water types and flow rates for both UV sources using combined variable modeling, which enabled us to estimate log inactivation of other microorganisms. Following UV disinfection, wastewater quality met the WHO standards for unrestricted irrigation.

scholarly journals Efficiency and Technological Reliability of Contaminant Removal in Household WWTPs with Activated Sludge

2021 ◽  
Vol 11 (4) ◽  
pp. 1889 ◽  
Author(s):  
Agnieszka Micek ◽  
Krzysztof Jóźwiakowski ◽  
Michał Marzec ◽  
Agnieszka Listosz ◽  
Tadeusz Grabowski
Keyword(s):  
Activated Sludge ◽  
Wastewater Treatment Plants ◽  
Oxygen Demand ◽  
Domestic Wastewater ◽  
Pollutant Removal ◽  
Treated Wastewater ◽  
Contaminant Removal ◽  
Nitrogen And Phosphorus ◽  
Wastewater Samples ◽  
Polish Law

The results of research on the efficiency and technological reliability of domestic wastewater purification in two household wastewater treatment plants (WWTPs) with activated sludge are presented in this paper. The studied facilities were located in the territory of the Roztocze National Park (Poland). The mean wastewater flow rate in the WWTPs was 1.0 and 1.6 m3/day. In 2017–2019, 20 series of analyses were done, and 40 wastewater samples were taken. On the basis of the received results, the efficiency of basic pollutant removal was determined. The technological reliability of the tested facilities was specified using the Weibull method. The average removal efficiencies for the biochemical oxygen demand in 5 days (BOD5) and chemical oxygen demand (COD) were 66–83% and 62–65%, respectively. Much lower effects were obtained for total suspended solids (TSS) and amounted to 17–48%, while the efficiency of total phosphorus (TP) and total nitrogen (TN) removal did not exceed 34%. The analyzed systems were characterized by the reliability of TSS, BOD5, and COD removal at the level of 76–96%. However, the reliability of TN and TP elimination was less than 5%. Thus, in the case of biogenic compounds, the analyzed systems did not guarantee that the quality of treated wastewater would meet the requirements of the Polish law during any period of operation. This disqualifies the discussed technological solution in terms of its wide application in protected areas and near lakes, where the requirements for nitrogen and phosphorus removal are high.

scholarly journals TIM-1 Promotes Japanese Encephalitis Virus Entry and Infection

Viruses ◽  
2018 ◽  
Vol 10 (11) ◽  
pp. 630 ◽  
Author(s):  
Jichen Niu ◽  
Ya Jiang ◽  
Hao Xu ◽  
Changjing Zhao ◽  
Guodong Zhou ◽  
...  
Keyword(s):  
Japanese Encephalitis Virus ◽  
Point Mutation ◽  
Japanese Encephalitis ◽  
Virus Entry ◽  
A549 Cells ◽  
Binding Pocket ◽  
Cytoplasmic Domain ◽  
Encephalitis Virus ◽  
Host Cells ◽  
Attachment Factor

Japanese encephalitis virus (JEV) is a mosquito-borne Flavivirus, the leading cause of viral-induced encephalitis. Several host molecules have been identified as the JEV attachment factor; however, the molecules involved in JEV entry remain poorly understood. In the present study, we demonstrate that TIM-1 is important for efficient infection by JEV. Firstly, three TIM-1 variants (V1, V2, and V3) were cloned from A549 cells, and we revealed that only ectopically TIM-1 V2 expression in 293T cells significantly promotes JEV attachment, entry and infection. Point mutation of phosphatidylserine (Ptdser) binding pocket in the TIM-1 IgV domain dampened JEV entry, indicating that TIM-1-mediated JEV infection is Ptdser-dependent. Furthermore, we found the cytoplasmic domain of TIM-1 is also required for enhancing JEV entry. Additionally, knock down of TIM-1 expression in A549 cells impaired JEV entry and infection, but not attachment, suggesting that additional factors exist in A549 cells that allow the virus to bind. In conclusion, our findings demonstrate that TIM-1 promotes JEV infection as an entry cofactor, and the polymorphism of TIM-1 is associated with JEV susceptibility to host cells.

scholarly journals AOA-2 Derivatives as Outer Membrane Protein A Inhibitors for Treatment of Gram-Negative Bacilli Infections

2021 ◽  
Vol 12 ◽  
Author(s):  
Rafael Ayerbe-Algaba ◽  
Nuria Bayó ◽  
Ester Verdú ◽  
Raquel Parra-Millán ◽  
Jesús Seco ◽  
...  
Keyword(s):  
Protein A ◽  
A549 Cells ◽  
Lung Epithelial Cells ◽  
Host Cells ◽  
Gram Negative ◽  
E Coli ◽  
Cyclic Hexapeptide ◽  
Diphenyltetrazolium Bromide ◽  
Peptide Derivatives

Previously, we identified that a cyclic hexapeptide AOA-2 inhibited the interaction of Gram-negative bacilli (GNB) like Acinetobacter baumannii, Pseudomonas aeruginosa, and Escherichia coli to host cells thereby preventing the development of infection in vitro and in a murine sepsis peritoneal model. In this work, we aimed to evaluate in vitro a library of AOA-2 derivatives in order to improve the effect of AOA-2 against GNB infections. Ten AOA-2 derivatives were synthetized for the in vitro assays. Their toxicities to human lung epithelial cells (A549 cells) for 24 h were evaluated by determining the A549 cells viability using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The effect of these peptide derivatives and AOA-2 at 250, 125, 62.5, and 31.25 μg/mL on the attachment of A. baumannii ATCC 17978, P. aeruginosa PAO1 and E. coli ATCC 25922 strains to A549 cells was characterized by adherence and viability assays. None of the 10 derivatives showed toxicity to A549 cells. RW01 and RW06 have reduced more the adherence of ATCC 17978, PAO1 and ATCC 2599 strains to A549 cells when compared with the original compound AOA-2. Moreover, both peptides have increased slightly the viability of infected A549 cells by PAO1 and ATCC 25922 than those observed with AOA-2. Finally, RW01 and RW06 have potentiated the activity of colistin against ATCC 17978 strain in the same level with AOA-2. The optimization program of AOA-2 has generated two derivatives (RW01 and RW06) with best effect against interaction of GNB with host cells, specifically against P. aeruginosa and E. coli.

scholarly journals LY6E Restricts Entry of Human Coronaviruses, Including Currently Pandemic SARS-CoV-2

2020 ◽  
Vol 94 (18) ◽  
Author(s):  
Xuesen Zhao ◽  
Shuangli Zheng ◽  
Danying Chen ◽  
Mei Zheng ◽  
Xinglin Li ◽  
...  
Keyword(s):  
Influenza A ◽  
Yellow Fever Virus ◽  
Virus Entry ◽  
Ectopic Expression ◽  
A549 Cells ◽  
Cellular Protein ◽  
Host Cells ◽  
Cellular Proteins ◽  
Human Coronavirus ◽  
Human Coronaviruses

ABSTRACT C3A is a subclone of the human hepatoblastoma HepG2 cell line with strong contact inhibition of growth. We fortuitously found that C3A was more susceptible to human coronavirus HCoV-OC43 infection than HepG2, which was attributed to the increased efficiency of virus entry into C3A cells. In an effort to search for the host cellular protein(s) mediating the differential susceptibility of the two cell lines to HCoV-OC43 infection, we found that ArfGAP with dual pleckstrin homology (PH) domains 2 (ADAP2), gamma-interferon-inducible lysosome/endosome-localized thiolreductase (GILT), and lymphocyte antigen 6 family member E (LY6E), the three cellular proteins identified to function in interference with virus entry, were expressed at significantly higher levels in HepG2 cells. Functional analyses revealed that ectopic expression of LY6E, but not GILT or ADAP2, in HEK 293 cells inhibited the entry of HCoV-O43. While overexpression of LY6E in C3A and A549 cells efficiently inhibited the infection of HCoV-OC43, knockdown of LY6E expression in HepG2 significantly increased its susceptibility to HCoV-OC43 infection. Moreover, we found that LY6E also efficiently restricted the entry mediated by the envelope spike proteins of other human coronaviruses, including the currently pandemic SARS-CoV-2. Interestingly, overexpression of serine protease TMPRSS2 or amphotericin treatment significantly neutralized the IFN-inducible transmembrane 3 (IFITM3) restriction of human coronavirus (CoV) entry, but did not compromise the effect of LY6E on the entry of human coronaviruses. The work reported herein thus demonstrates that LY6E is a critical antiviral immune effector that controls CoV infection and pathogenesis via a mechanism distinct from other factors that modulate CoV entry. IMPORTANCE Virus entry into host cells is one of the key determinants of host range and cell tropism and is subjected to the control of host innate and adaptive immune responses. In the last decade, several interferon-inducible cellular proteins, including IFITMs, GILT, ADAP2, 25CH, and LY6E, had been identified to modulate the infectious entry of a variety of viruses. Particularly, LY6E was recently identified as a host factor that facilitates the entry of several human-pathogenic viruses, including human immunodeficiency virus, influenza A virus, and yellow fever virus. Identification of LY6E as a potent restriction factor of coronaviruses expands the biological function of LY6E and sheds new light on the immunopathogenesis of human coronavirus infection.

scholarly journals Intracellular Trafficking and Persistence of Acinetobacter baumannii Requires Transcription Factor EB

mSphere ◽  
2018 ◽  
Vol 3 (2) ◽  
Author(s):  
Raquel Parra-Millán ◽  
David Guerrero-Gómez ◽  
Rafael Ayerbe-Algaba ◽  
Maria Eugenia Pachón-Ibáñez ◽  
Antonio Miranda-Vizuete ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Acinetobacter Baumannii ◽  
Intracellular Trafficking ◽  
Nuclear Translocation ◽  
A549 Cells ◽  
Lung Epithelial Cells ◽  
Host Cells ◽  
Infection Model ◽  
Content Type ◽  
Transcription Factor Eb

ABSTRACT Acinetobacter baumannii is a significant human pathogen associated with hospital-acquired infections. While adhesion, an initial and important step in A. baumannii infection, is well characterized, the intracellular trafficking of this pathogen inside host cells remains poorly studied. Here, we demonstrate that transcription factor EB (TFEB) is activated after A. baumannii infection of human lung epithelial cells (A549). We also show that TFEB is required for the invasion and persistence inside A549 cells. Consequently, lysosomal biogenesis and autophagy activation were observed after TFEB activation which could increase the death of A549 cells. In addition, using the Caenorhabditis elegans infection model by A. baumannii , the TFEB orthologue HLH-30 was required for survival of the nematode to infection, although nuclear translocation of HLH-30 was not required. These results identify TFEB as a conserved key factor in the pathogenesis of A. baumannii . IMPORTANCE Adhesion is an initial and important step in Acinetobacter baumannii infections. However, the mechanism of entrance and persistence inside host cells is unclear and remains to be understood. In this study, we report that, in addition to its known role in host defense against Gram-positive bacterial infection, TFEB also plays an important role in the intracellular trafficking of A. baumannii in host cells. TFEB was activated shortly after A. baumannii infection and is required for its persistence within host cells. Additionally, using the C. elegans infection model by A. baumannii , the TFEB orthologue HLH-30 was required for survival of the nematode to infection, although nuclear translocation of HLH-30 was not required.

scholarly journals Effect of Host Cells on Low- and Medium-Pressure UV Inactivation of Adenoviruses

2010 ◽  
Vol 76 (21) ◽  
pp. 7068-7075 ◽  
Author(s):  
Huiling Guo ◽  
Xiaona Chu ◽  
Jiangyong Hu
Keyword(s):  
Host Cell ◽  
Cell Line ◽  
Cell Lines ◽  
Host Cells ◽  
Uv Disinfection ◽  
Human Embryonic Kidney ◽  
Experimental Conditions ◽  
Medium Pressure ◽  
Serotype 5 ◽  
Uv Dose

ABSTRACT UV disinfection is highly effective against most pathogens, with the exception of the adenoviruses (AD). To date, many studies have focused on low-pressure (LP) UV inactivation of AD, but little is known about the effect of medium-pressure (MP) UV inactivation of AD. Despite numerous studies of LP UV inactivation of AD, extreme variabilities in the LP UV dose requirements of AD had been observed because of differing experimental conditions used, such as the types of cell lines used for AD enumeration. This study therefore investigates the effect of three different host cell lines (PLC/PRF/5, human embryonic kidney 293 [HEK293], and XP17BE) on the LP and MP UV dose requirements of AD serotype 5 (AD5), AD40, and AD41 under similar experimental settings. Results showed that for 4-log inactivation of AD, LP UV and MP UV doses needed to be in the ranges of 123 to 182 mJ/cm2 and 65 to 90 mJ/cm2, respectively, when HEK293 and PLC/PRF/5 cells were used for enumeration. The UV doses required for MP UV inactivation of AD were significantly lower than those required for LP UV inactivation (P value < 0.05). When different cell lines were used for enumeration, UV dose requirements for AD differed. AD were portrayed to be most susceptible to UV (LP UV doses of <57 mJ/cm2 and MP UV doses of <42 mJ/cm2 for 4-log AD inactivation) when the XP17BE cells were used as the host cell. The use of different cell lines for AD enumeration affected LP UV dose results more significantly than MP UV dose results (P value < 0.05). Cell line variability factors for LP UV disinfection (CLLP) and MP UV disinfection (CLMP) for AD5, AD40, and AD41 enumerated with HEK293, PLC/PRF/5, and XP17BE cells were in the ranges of 1.0 to 3.2 and 1.0 to 2.5, respectively.

Toxicity Screening Tests for Wastewater Treatment Plants – I

2005 ◽  
Vol 277-279 ◽  
pp. 559-568
Author(s):  
Jin Hwa Park ◽  
Yong Woo Yl
Keyword(s):  
Electron Transfer ◽  
Wastewater Treatment Plants ◽  
Domestic Wastewater ◽  
Strong Relationship ◽  
Domestic Sewage ◽  
Screening Tests ◽  
Toxicity Screening ◽  
Trickling Filter ◽  
Home Life ◽  
Wastewater Samples

Toxicity screening tests using the Reserve Electron Transfer (RET) and Electron Transfer (ETr) assays were performed with five wastewater samples amended with trickling filter (TF) or activated sludge (AS) biomass. In the case of untreated samples, Home Life domestic wastewater (HLD/W) showed the lowest inhibition, followed by domestic sewage (DS), hospital wastewater (H/W), East Straus wastewater (ES/W), and Mills wastewater (M/W) from both ETr and RET assays. After 12 hours of treatment at 20°C, DS with AS biomass had the lowest % inhibition from the RET assay, followed by DS with TF, HLD/W with TF, H/W with TF, M/W with TF, and M/W with AS. AS biomass reduced more toxicity from DS than TF biomass whereas acclimated TF biomass reduced significantly more toxicity than AS biomass, indicating the importance of acclimation. M/W was most toxic and resistant to biodegradation among six wastewater samples. No nitrification occurred with M/W and ES/W. While there was significant nitrification with DS treated by AS biomass, little nitrification by TF biomass occurred even with DS, HLD/W, and H/W. It appears that nitrification is significantly inhibited by M/W and ES/W even when mixed with domestic sewage. It appears that there is a strong relationship between the TIC/TC ratio and % inhibition.

scholarly journals iTRAQ-Based and Label-Free Proteomics Approaches for Studies of Human Adenovirus Infections

2013 ◽  
Vol 2013 ◽  
pp. 1-16 ◽  
Author(s):  
Hung V. Trinh ◽  
Jonas Grossmann ◽  
Peter Gehrig ◽  
Bernd Roschitzki ◽  
Ralph Schlapbach ◽  
...  
Keyword(s):  
A549 Cells ◽  
Correlation Coefficients ◽  
Human Adenovirus ◽  
Adenovirus Type ◽  
Label Free ◽  
Absolute Quantitation ◽  
Software Packages ◽  
Itraq Label ◽  
Isobaric Tags ◽  
Type 3

Both isobaric tags for relative and absolute quantitation (iTRAQ) and label-free methods are widely used for quantitative proteomics. Here, we provide a detailed evaluation of these proteomics approaches based on large datasets from biological samples. iTRAQ-label-based and label-free quantitations were compared using protein lysate samples from noninfected human lung epithelial A549 cells and from cells infected for 24 h with human adenovirus type 3 or type 5. Either iTRAQ-label-based or label-free methods were used, and the resulting samples were analyzed by liquid chromatography (LC) and tandem mass spectrometry (MS/MS). To reduce a possible bias from quantitation software, we applied several software packages for each procedure. ProteinPilot and Scaffold Q+ software were used for iTRAQ-labeled samples, while Progenesis LC-MS and ProgenesisF-T2PQ/T3PQ were employed for label-free analyses. R2 correlation coefficients correlated well between two software packages applied to the same datasets with values between 0.48 and 0.78 for iTRAQ-label-based quantitations and 0.5 and 0.86 for label-free quantitations. Analyses of label-free samples showed higher levels of protein up- or downregulation in comparison to iTRAQ-labeled samples. The concentration differences were further evaluated by Western blotting for four downregulated proteins. These data suggested that the label-free method was more accurate than the iTRAQ method.

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