Effect of Nk-lysin peptides on bacterial growth, MIC, antimicrobial resistance, and viral activities.

NK-lysins from chicken, bovine and human are used as antiviral and antibacterial agents. Gram-negative and gram-positive microorganisms, including Streptococcus pyogenes, Streptococcus mutans, Escherichia coli, Pseudomonas aeruginosa, Klebsiella oxytoca, Shigella sonnei, Klebsiella pneumoniae and Salmonella typhimurium, are susceptible to NK-lysin treatment. The presence of dominant TEM-1 gene was noted in all untreated and treated bacteria, while TOHO-1 gene was absent in all bacteria. Importantly, ß-lactamase genes CTX-M-1, CTX-M-8, and CTX-M-9 genes were detected in untreated bacterial strains; however, none of these were found in any bacterial strains following treatment with NK-lysin peptides. NK-lysin peptides are also used to test for inhibition of infectivity, which ranged from 50 to 90% depending on NK-lysin species. Chicken, bo vine and human NK-lysin peptides are demonstrated herein to have antibacterial activity and antiviral activity against Rotavirus (strain SA-11). On the basis of the comparison between these peptides, potent antiviral activity of bovine NK-lysin against Rotavirus (strain SA-11) is particularly evident, inhibiting infection by up to 90%. However, growth was also significantly inhibited by chicken and human NK-lysin peptides, restricted by 80 and 50%, respectively. This study provided a novel treatment using NK-lysin peptides to inhibit expression of ß-lactamase genes in ß-lactam antibiotic-resistant bacterial infections.

Modeling the role of fomites in a norovirus outbreak.

Norovirus accounts for a large portion of the gastroenteritis disease burden, and outbreaks have occurred in a wide variety of environments. Understanding the role of fomites in norovirus transmission will inform behavioral interventions, such as hand washing and surface disinfection. The purpose of this study was to estimate the contribution of fomite-mediated exposures to infection and illness risks in outbreaks. A simulation model in discrete time that accounted for hand-to-porous surfaces, hand-to-nonporous surfaces, hand-to-mouth, -eyes, -nose, and hand washing events was used to predict 17 hr of simulated human behavior. Norovirus concentrations originated from monitoring contamination levels on surfaces during an outbreak on houseboats. To predict infection risk, two dose-response models (fractional Poisson and 2F1 hypergeometric) were used to capture a range of infection risks. A triangular distribution describing the conditional probability of illness given an infection was multiplied by modeled infection risks to estimate illness risks. Infection risks ranged from 70.22% to 72.20% and illness risks ranged from 21.29% to 70.36%. A sensitivity analysis revealed that the number of hand-to-mouth contacts and the number of hand washing events had strong relationships with model-predicted doses. Predicted illness risks overlapped with leisure setting and environmental attack rates reported in the literature. In the outbreak associated with the viral concentrations used in this study, attack rates ranged from 50% to 86%. This model suggests that fomites may have accounted for 25% to 82% of illnesses in this outbreak. Fomite-mediated exposures may contribute to a large portion of total attack rates in outbreaks involving multiple transmission modes. The findings of this study reinforce the importance of frequent fomite cleaning and hand washing, especially when ill persons are present.

Applicability of crAssphage as a performance indicator for viral reduction during activated sludge wastewater treatment.

A major threat to water quality is the discharge of human-derived wastewater, which can cause waterborne illnesses associated with enteric viruses. A poor association exists between fecal indicator bacteria and virus fate in the environment, especially during wastewater treatment. In the current study, the potential of using a novel human gut bacteriophage crAssphage as a wastewater treatment process indicator was evaluated. Using qPCR, influent and effluent wastewater samples of two wastewater treatment plants were analyzed for crAssphage and human viruses including human bocavirus (HBoV), human adenovirus (HAdV), and human polyomavirus (HPyV). All samples were positive for crAssphage. The annual crAssphage concentrations varied between 1.45E + 04 and 2.39E + 08 gc/l in influent samples and from 1.25E + 04 to 7.88E + 06 gc/l in effluent samples. Human viruses concentrations were some orders of magnitude lower than that of crAssphage. Data demonstrated a significant correlation between crAssphage, HAdV, and HPyV during the wastewater treatment process, suggesting that crAssphage and human viral pathogens have similar removal mechanisms. Ultimately, this work concludes that crAssphage could be a performance indicator for viral reduction in the wastewater treatment process.

Cold atmospheric plasma: a sustainable approach to inactivating viruses, bacteria, and protozoa with remediation of organic pollutants in river water and wastewater.

Innovative technologies are needed to enhance access to clean water and avoid waterborne diseases. We investigated the performance of cold atmospheric plasma (CAP), a clean and sustainable approach for microbial inactivation and total organic carbon (TOC) degradation in environmental water. Water matrices played a crucial role in the performance of CAP efficacy; for example, complete removal of ɸX174 from dH2O required 1 min of treatment, while ɸX174 reductions of ~ 2log10 and 4log10 were obtained after 10 min of CAP exposure in river water and wastewater samples, respectively. Similarly, after 10 min of CAP treatment, bacterial concentrations decreased by 3 log10 and 4 log10, in river and wastewater samples, respectively. In contrast, after 30 s of contact time, a 4 log10 reduction of bacteria was accomplished in dH2O. Complete removal of Acanthamoeba from dH2O was found after 30 min of CAP treatment, whereas it was not removed from surface water or wastewater at the same exposure time. Additionally, the approach successfully reduced TOC, and the degradation kinetics of TOC were represented by pseudo-first-order. CAP showed higher rates of TOC degradation in the final effluent of the wastewater treatment plant compared to surface water. The difference in CAP performance between river water and wastewater could be attributed to the bulk structure of humic acids in river water compared to small organic byproducts in the final effluent of WWTP. Overall, the findings reported here support the idea that CAP holds promise as a sustainable solution for controlling pathogens, removing organic water pollution, and integrating with traditional purification processes. Low-cost systems may advance CAP technology and increase its widespread use.

Removal of chemical and microbial water pollutants by cold plasma combined with Ag/TiO2-rGO nanoparticles.

This study aimed to investigate the synergistic effect of the cold atmospheric plasma (CAP) and heterogeneous photocatalytic processes in an aqueous solution to enhance water purification efficacy and reduce the energy cost required by CAP. 0.1% Ag/TiO2-reduced graphene oxide (rGO) nanoparticles (NPs) photo-composite were prepared and fully characterized. Data showed that Ag nanoparticles and the rGO play an important role in increasing the efficiency of the whole treatment process and the photo-composite (0.1% Ag/TiO2-1% rGO at 400 °C) revealed the highest phenol removal rate with excellent reusability. Also, complete inactivation (~ 5log10 reduction) of both E. coli and S. aureus by NPs was observed without CAP exposure, whereas a minimal effect (0.1-0.5 log10) on viruses (Adenovirus (AdV), rotavirus, and ɸX174) was observed after 10 min incubation. Interestingly, the photocatalytic virus inactivation test was promising, as it resulted in > 4.7log10 reduction of AdV at 2 min treatment, whereas < 1log10 could be reduced using only CAP at the same treatment time. Accordingly, we believe that this work could provide new insights into how the synergy between CAP and 0.1% Ag/TiO2-1% rGO photo-composite in aqueous media imposes a great potential for environmental applications, such as water purification and microbial inactivation.

Etiology of diarrheal disease among children under 5 years in Egypt: a high incidence of human bocavirus.

BACKGROUND: Human bocavirus (HBoV) is globally distributed and associated with respiratory and enteric infections. Limited data are available about the incidence of HBoV in Egyptian children. We aimed to investigate the association of HBoV genotypes in children with diarrheal disease and also to determine the possibility of HBoV co-infections with other human enteric pathogens. METHODS: A total of 102 stool samples were collected from children under five years old with diarrhea. Samples were analyzed for the presence of HBoV by real-time PCR. HBoV positive samples were tested for adenovirus (AdV), rotavirus (RoV), parasitic helminths, and enteric protozoa. RESULTS: HBoV was detected in 58% of examined cases. HBoV-3 was the most prevalent genotype observed (44%; 45 of 102), followed by HBoV-2/4 (33%; 34 of 102) and HBoV-1 (30%; 31 of 102). Although the incidence of HBoV was higher in males (66.6%; 34 of 51) than females (49%; 25 of 51), the analysis showed no significant difference for HBoV between genders. The average HBoV concentrations were 5.3 × 104 GC/g in males and 1.03 × 105 GC/g in females. Among the HBoV-positive samples, the single infection of HBoV was 52.5% (31/59), while the co-infections with multiple viruses were found in 1.7% (1/59) for HBoV and AdV, 33.9% (20/59) for HBoV and RoV, and 11.9% (7/59) for HBoV, and RoV and AdV. No co-infection with parasitic helminths or enteric protozoa was found. CONCLUSIONS: The single infection of HBoV in some children suffering from acute gastroenteritis indicated that HBoV could be the main etiologic agent of the disease. The study highlights the high incidence of HBoVs genotypes with remarkable multiple co-infections in the pre-school children in Egypt.

Comparative Assessment of BGM and PLC/PRF/5 Cell Lines for Enteric Virus Detection in Biosolids.

The buffalo green monkey (BGM) cell line is required for the detection of enteric viruses in biosolids through a total culturable viral assay (TCVA) by the United States Environmental Protection Agency. In the present study, BGM and PLC/PRF/5 cell lines were evaluated for TCVA and for their use in determining the incidence of adenoviruses and enteroviruses in raw sludge and Class B biosolids. Six raw sludge and 17 Class B biosolid samples were collected from 13 wastewater treatment plants from seven U.S. states. Samples were processed via organic flocculation and concentrate volumes equivalent to 4 g total solids were assayed on BGM and PLC/PRF/5 cells. Cell monolayers were observed for cytopathic effect (CPE) after two 14-days passages. Cell lysates were tested for the presence of adenoviruses and enteroviruses by PCR or RT-PCR. The PLC/PRF/5 cells detected more culturable viruses than the BGM cells by CPE (73.9% vs. 56.5%, respectively). 52% of the samples were positive for CPE using both cell lines. No viruses were detected in either cell line by PCR in flasks in which CPE was not observed. No adenoviruses were detected in 13 CPE-positive samples from BGM lysates. In contrast, of the 17 samples exhibiting CPE on PLC/PRF/5 cells, 14 were positive for adenoviruses (82.4%). In conclusion, PLC/PRF/5 cells were superior for the detection of adenoviruses in both raw sludge and Class B biosolids. Thus, the use of BGM cells alone for TCVA may underestimate the viral concentration in sludge/biosolid samples.

Efficiency of Reovirus Concentration from Water with Positively Charged Filters.

This study examined the efficacy of reovirus concentration from large volumes of water using two positively charged filters: Zeta Plus 1MDS and NanoCeram. The results indicated that an average of 61 and 81% of input reoviruses were effectively recovered, respectively, from recycled water and tap water using NanoCeram filtration.

Comparative survival of viruses during thermophilic and mesophilic anaerobic digestion.

Micro-scale technology was used to obtain survival data for three animal viruses and two bacteriophages during anaerobic digestion. The data for adenoviruses, MS2 and Φ6 provide the first published reports for survival of these viruses during mesophilic anaerobic digestion. Data were also obtained for thermophilic digestion, which showed greater inactivation of viruses at the higher temperature. Data on the survival of Φ6 are of particular interest since it is a lipid-based virus which has been suggested as a surrogate for the Ebola virus. In contrast, MS2 was found to be an inappropriate surrogate for Ebola.

Wide-spectrum activity of a silver-impregnated fabric.

Fabrics, such as clothing, drapes, pillowcases, and bedsheets are potential sources of pathogenic bacteria and viruses. We found fabrics (ie, professional clothing, pillowcases, and lab coats) treated with a silver-impregnated material to be effective in significantly reducing a wide spectrum of ordinary and drug-resistant microorganisms, including Salmonella, methicillin-resistant Staphylococcus aureus, Propionibacterium acnes, Trichphyton mentagrophytes, and norovirus. Fabrics impregnated with antimicrobial agents help provide an additional barrier to the transport or reservoir of pathogens in health care environments.

Assessment of coliphage surrogates for testing drinking water treatment devices.

Test protocols have been developed by the United States Environmental Protection Agency (USEPA) and the World Health Organization (WHO) to test water treatment devices/systems that are used at the individual and home levels to ensure the removal of waterborne viruses. The goal of this study was to assess if coliphage surrogates could be used in this testing in place of the currently required use of animal or human enteric viruses. Five different coliphages (MS-2, PRD1, ΦX-174, Qß, and fr) were compared to the removal of poliovirus type 1 (LSc-2ab) by eight different water treatment devices/systems using a general case and a challenge case (high organic load, dissolved solids, and turbidity) test water as defined by the USEPA. The performance of the units was rated as a pass/fail based on a 4 log removal/inactivation of the viruses. In all cases, a failure or a pass of the units/system for poliovirus also corresponded to a pass/fail by all of the coliphages. In summary, in using pass/fail criteria as recommended under USEPA guidelines for testing water treatment device/systems, the use of coliphages should be considered as an alternative to reduce cost and time of testing such devices/systems.

Simultaneous Concentration of Bovine Viruses and Agricultural Zoonotic Bacteria from Water Using Sodocalcic Glass Wool Filters.

Infiltration and runoff from manured agricultural fields can result in livestock pathogens reaching groundwater and surface waters. Here, we measured the effectiveness of glass wool filters to simultaneously concentrate enteric viruses and bacteria of bovine origin from water. The recovery efficiencies were determined for bovine viral diarrhea virus types 1 and 2, bovine rotavirus group A, bovine coronavirus, poliovirus Sabin III, toxigenic Escherichia coli ,and Campylobacter jejuni seeded into water with three different turbidity levels (0.5, 215, and 447 NTU). Twenty liters of dechlorinated tap water (pH 7) were seeded with the test organisms, and then passed through a glass wool filter using a peristaltic pump (flow rate = 1 liter min(-1)). Retained organisms were eluted from the filters by passing beef extract-glycine buffer (pH 9.5) in the direction opposite of sample flow. Recovered organisms were enumerated by qPCR except for C. jejuni, which was quantified by culture. Mean recovery efficiencies ranged from 55 to 33% for the bacteria and 58 to 16% for the viruses. Using bootstrapping techniques combined with Analysis of Variance, recovery efficiencies were found to differ among the pathogen types tested at the two lowest turbidity levels; however, for a given pathogen type turbidity did not affect recovery except for C. jejuni. Glass wool filtration is a cost-effective method for concentrating several waterborne pathogens of bovine origin simultaneously, although recovery may be low for some specific taxa such as bovine viral diarrhea virus 1.

Assessment of a portable handheld UV light device for the disinfection of viruses and bacteria in water.

Effective individual microbiological water purifiers are needed for consumption of untreated water sources by campers, emergency use, military, and in developing counties. A handheld UV light device was tested to assess if it could meet the virus reduction requirements established by the United State Environmental Protection Agency, National Science Foundation and the World Health Organization. The device was found capable of inactivating at least 4 log10 of poliovirus type 1, rotavirus SA-11 and MS-2 virus in 500 mL volumes of general case test water. But in the presence of high turbidity and organic matter, filtration was necessary to achieve a 4 log10 reduction of the test viruses.