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1.
Arch Virol ; 169(7): 142, 2024 Jun 08.
Article in English | MEDLINE | ID: mdl-38851653

ABSTRACT

Uropathogenic Escherichia coli (UPEC) is the most common causative agent of urinary tract infections, and strains that are resistant to antibiotics are a major problem in treating these infections. Phage therapy is a promising alternative approach that can be used to treat infections caused by polyresistant bacterial strains. In the present study, 16 bacteriophages isolated from sewage and surface water were investigated. Phage host specificity was tested on a collection of 77 UPEC strains. The phages infected 2-44 strains, and 80% of the strains were infected by at least one phage. The susceptible E. coli strains belonged predominantly to the B2 phylogenetic group, including strains of two clones, CC131 and CC73, that have a worldwide distribution. All of the phages belonged to class Caudoviricetes and were identified as members of the families Straboviridae, Autographiviridae, and Drexlerviridae and the genera Kagunavirus, Justusliebigvirus, and Murrayvirus. A phage cocktail composed of six phages - four members of the family Straboviridae and two members of the family Autographiviridae - was prepared, and its antibacterial activity was tested in liquid medium. Complete suppression of bacterial growth was observed after 5-22 hours of cultivation, followed by partial regrowth. At 24 hours postinfection, the cocktail suppressed bacterial growth to 43-92% of control values. Similar results were obtained when testing the activity of the phage cocktail in LB and in artificial urine medium. The results indicate that our phage cocktail has potential to inhibit bacterial growth during infection, and they will therefore be preserved in the national phage bank, serving as valuable resources for therapeutic applications.


Subject(s)
Drug Resistance, Multiple, Bacterial , Host Specificity , Phylogeny , Uropathogenic Escherichia coli , Uropathogenic Escherichia coli/virology , Uropathogenic Escherichia coli/drug effects , Bacteriophages/classification , Bacteriophages/physiology , Bacteriophages/genetics , Bacteriophages/isolation & purification , Sewage/virology , Phage Therapy/methods , Humans , Anti-Bacterial Agents/pharmacology , Escherichia coli Infections/microbiology , Escherichia coli Infections/therapy
2.
Front Cell Infect Microbiol ; 14: 1382145, 2024.
Article in English | MEDLINE | ID: mdl-38736748

ABSTRACT

Carbapenem-resistant Acinetobacter baumannii (CRAB) has become a new threat in recent years, owing to its rapidly increasing resistance to antibiotics and new effective therapies are needed to combat this pathogen. Phage therapy is considered to be the most promising alternative for treating CRAB infections. In this study, a novel phage, Ab_WF01, which can lyse clinical CRAB, was isolated and characterized from hospital sewage. The multiplicity of infection, morphology, one-step growth curve, stability, sensitivity, and lytic activity of the phage were also investigated. The genome of phage Ab_WF01 was 41, 317 bp in size with a GC content of 39.12% and encoded 51 open reading frames (ORFs). tRNA, virulence, and antibiotic resistance genes were not detected in the phage genome. Comparative genomic and phylogenetic analyses suggest that phage Ab_WF01 is a novel species of the genus Friunavirus, subfamily Beijerinckvirinae, and family Autographiviridae. The in vivo results showed that phage Ab_WF01 significantly increased the survival rate of CRAB-infected Galleria mellonella (from 0% to 70% at 48 h) and mice (from 0% to 60% for 7 days). Moreover, after day 3 post-infection, phage Ab_WF01 reduced inflammatory response, with strongly ameliorated histological damage and bacterial clearance in infected tissue organs (lungs, liver, and spleen) in mouse CRAB infection model. Taken together, these results show that phage Ab_WF01 holds great promise as a potential alternative agent with excellent stability for against CRAB infections.


Subject(s)
Acinetobacter Infections , Acinetobacter baumannii , Bacteriophages , Carbapenems , Genome, Viral , Phage Therapy , Phylogeny , Sewage , Acinetobacter baumannii/virology , Acinetobacter baumannii/drug effects , Sewage/virology , Sewage/microbiology , Animals , Carbapenems/pharmacology , Bacteriophages/genetics , Bacteriophages/physiology , Bacteriophages/classification , Bacteriophages/isolation & purification , Acinetobacter Infections/microbiology , Mice , Anti-Bacterial Agents/pharmacology , Open Reading Frames , Disease Models, Animal , Moths/virology , Moths/microbiology , Base Composition
3.
PLoS One ; 19(5): e0302000, 2024.
Article in English | MEDLINE | ID: mdl-38709720

ABSTRACT

Wastewater surveillance represents an alternative approach to regulating contamination and the early detection of infectious agents and outbreaks of diseases of public health importance. This study evaluated domestic wastewater effects on recreational waters in estuarine and seawater bodies in Guayas and Santa Elena provinces in Ecuador, South America. Fecal indicator bacteria (thermotolerant coliforms) served as key indicators for evaluation. Physical, chemical, and microbiological quality markers following the Ecuadorian environmental quality standard and the discharge of effluents to the water resource were analyzed. Samples were collected from 44 coastal sites and 2 oxidation lagoons during the dry and rainy seasons of 2020 and 2021, respectively. SARS-CoV-2 RNA was detected in samples with higher E. coli concentrations using reverse transcription quantitative PCR to detect the genes N and ORF1ab. All samples analyzed for SARS-CoV-2 showed Ct ˂ 40 for at least one gene. Four samples showed at least 20 genome copies of gene N per reaction. These were at an artisanal fishing port, an estuarine area (Palmar), a recreational bay, and an oxidation lagoon. A moderate correlation was found between SARS-CoV-2 RNA, thermotolerant coliform and E. coli (p-value ≤ 0.0037), and a strong and positive correlation between thermotolerant coliform and E. coli. (p-value ≤ 0.00001), highlighting the utility of these established parameters as a proxy of the virus. Significant differences were found in the concentrations of thermotolerant coliforms between seasons (p-value = 0.016) and sites (p-value = 0.005). The highest levels of coliforms were found in the dry season (63000 MPN/100 mL) in Anconcito and during the rainy season (14000 MPN/100 mL) at Esterillo in Playas County. It is recommended that the decentralized autonomous governments of the surveyed provinces in Ecuador implement urgent corrective actions and establish medium-term mechanisms to minimize a potential contamination route. Additional parameters must be included in the monitoring, such as Enterococcus and intestinal parasites, due to their public health implications. In the oxidation lagoons, maintenance actions must be carried out, including the dissolution of sediments, an increase in water retention times, and in situ treatment of the sludge, to improve the system's performance.


Subject(s)
COVID-19 , RNA, Viral , SARS-CoV-2 , Sewage , Water Quality , Ecuador , Sewage/virology , Sewage/microbiology , SARS-CoV-2/isolation & purification , SARS-CoV-2/genetics , RNA, Viral/genetics , RNA, Viral/isolation & purification , RNA, Viral/analysis , COVID-19/epidemiology , COVID-19/virology , Humans , Enterobacteriaceae/isolation & purification , Enterobacteriaceae/genetics , Water Microbiology , Environmental Monitoring/methods , Seawater/virology , Seawater/microbiology , Escherichia coli/genetics , Escherichia coli/isolation & purification , Wastewater/virology , Wastewater/microbiology
4.
PLoS One ; 19(5): e0303529, 2024.
Article in English | MEDLINE | ID: mdl-38809825

ABSTRACT

Wastewater-based epidemiology (WBE) has emerged as an effective environmental surveillance tool for predicting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) disease outbreaks in high-income countries (HICs) with centralized sewage infrastructure. However, few studies have applied WBE alongside epidemic disease modelling to estimate the prevalence of SARS-CoV-2 in low-resource settings. This study aimed to explore the feasibility of collecting untreated wastewater samples from rural and urban catchment areas of Nagpur district, to detect and quantify SARS-CoV-2 using real-time qPCR, to compare geographic differences in viral loads, and to integrate the wastewater data into a modified Susceptible-Exposed-Infectious-Confirmed Positives-Recovered (SEIPR) model. Of the 983 wastewater samples analyzed for SARS-CoV-2 RNA, we detected significantly higher sample positivity rates, 43.7% (95% confidence interval (CI) 40.1, 47.4) and 30.4% (95% CI 24.66, 36.66), and higher viral loads for the urban compared with rural samples, respectively. The Basic reproductive number, R0, positively correlated with population density and negatively correlated with humidity, a proxy for rainfall and dilution of waste in the sewers. The SEIPR model estimated the rate of unreported coronavirus disease 2019 (COVID-19) cases at the start of the wave as 13.97 [95% CI (10.17, 17.0)] times that of confirmed cases, representing a material difference in cases and healthcare resource burden. Wastewater surveillance might prove to be a more reliable way to prepare for surges in COVID-19 cases during future waves for authorities.


Subject(s)
COVID-19 , SARS-CoV-2 , Wastewater , India/epidemiology , COVID-19/epidemiology , COVID-19/virology , COVID-19/diagnosis , Humans , Wastewater/virology , SARS-CoV-2/isolation & purification , Viral Load , Pandemics , Wastewater-Based Epidemiological Monitoring , Sewage/virology
5.
Vet Microbiol ; 294: 110133, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38820726

ABSTRACT

Non-aureus staphylococci (NAS) are an essential group of bacteria causing antimicrobial resistant intramammary infections in livestock, particularly dairy cows. Therefore, bacteriophages emerge as a potent bactericidal agent for NAS mastitis. This study aimed to obtain NAS-specific bacteriophages using bacterial strains isolated from cows with mastitis, subsequently evaluating their morphological, genomic, and lytic characteristics. Four distinct NAS bacteriophages were recovered from sewage or the environment of Chinese dairy farms; PT1-1, PT94, and PT1-9 were isolated using Staphylococcus chromogenes and PT1-4 using Staphylococcus gallinarum. Both PT1-1 (24/54, 44 %) and PT94 (28/54, 52 %) had broader lysis than PT1-4 (3/54, 6 %) and PT1-9 (10/54, 19 %), but PT1-4 and PT1-9 achieved cross-species lysis. All bacteriophages had a short latency period and good environmental tolerance, including surviving at pH=4-10 and at 30-60℃. Except for PT1-9, all bacteriophages had excellent bactericidal efficacy within 5 h of co-culture with host bacteria in vitro at various multiplicity of infection (MOIs). Based on whole genome sequencing, average nucleotide identity (ANI) analysis of PT1-1 and PT94 can be classified as the same species, consistent with whole-genome synteny analysis. Although motifs shared by the 4 bacteriophages differed little from those of other bacteriophages, a phylogenetic tree based on functional proteins indicated their novelty. Moreover, based on whole genome comparisons, we inferred that cross-species lysis of bacteriophage may be related to the presence of "phage tail fiber." In conclusion 4 novel NAS bacteriophages were isolated; they had good biological properties and unique genomes, with potential for NAS mastitis therapy.


Subject(s)
Genome, Viral , Mastitis, Bovine , Sewage , Staphylococcus , Sewage/virology , Sewage/microbiology , Animals , Staphylococcus/virology , Staphylococcus/drug effects , Staphylococcus/genetics , Cattle , Female , Mastitis, Bovine/microbiology , Staphylococcus Phages/genetics , Staphylococcus Phages/physiology , Staphylococcus Phages/classification , Bacteriophages/genetics , Bacteriophages/isolation & purification , Bacteriophages/classification , Bacteriophages/physiology , Staphylococcal Infections/veterinary , Staphylococcal Infections/microbiology , Phylogeny , Genomics , Whole Genome Sequencing
6.
Virology ; 595: 110098, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38705084

ABSTRACT

Acinetobacter baumannii is one of the most important pathogens of healthcare-associated infections. The rising prevalence of multidrug-resistant A. baumannii (MRAB) strains and biofilm formation impact the outcome of conventional treatment. Phage-related therapy is a promising strategy to tame troublesome multidrug-resistant bacteria. Here, we isolated and evaluated a highly efficient lytic phage called MRABP9 from hospital sewage. The phage was a novel species within the genus Friunavirus and exhibited lytic activity against 2 other identified MRAB strains. Genomic analysis revealed it was a safe virulent phage and a pectate lyase domain was identified within its tail spike protein. MRABP9 showed potent bactericidal and anti-biofilm activity against MRAB, significantly delaying the time point of bacterial regrowth in vitro. Phage administration could rescue the mice from acute lethal MRAB infection. Considering its features, MRABP9 has the potential as an efficient candidate for prophylactic and therapeutic use against acute infections caused by MRAB strains.


Subject(s)
Acinetobacter Infections , Acinetobacter baumannii , Bacteriophages , Drug Resistance, Multiple, Bacterial , Phage Therapy , Acinetobacter baumannii/virology , Acinetobacter baumannii/drug effects , Animals , Acinetobacter Infections/microbiology , Acinetobacter Infections/therapy , Mice , Bacteriophages/genetics , Bacteriophages/physiology , Phage Therapy/methods , Genome, Viral , Biofilms/drug effects , Biofilms/growth & development , Humans , Female , Sewage/virology
7.
Viruses ; 16(4)2024 03 29.
Article in English | MEDLINE | ID: mdl-38675877

ABSTRACT

The concentration of viruses in sewage sludge is significantly higher (10-1000-fold) than that found in natural environments, posing a potential risk for human and animal health. However, the composition of these viruses and their role in the transfer of pathogenic factors, as well as their role in the carbon, nitrogen, and phosphorus cycles remain poorly understood. In this study, we employed a shotgun metagenomic approach to investigate the pathogenic bacteria and viral composition and function in two wastewater treatment plants located on a campus. Our analysis revealed the presence of 1334 amplicon sequence variants (ASVs) across six sludge samples, with 242 ASVs (41.22% of total reads) identified as pathogenic bacteria. Arcobacter was found to be the most dominant pathogen accounting for 6.79% of total reads. The virome analysis identified 613 viral genera with Aorunvirus being the most abundant genus at 41.85%. Approximately 0.66% of these viruses were associated with human and animal diseases. More than 60% of the virome consisted of lytic phages. Host prediction analysis revealed that the phages primarily infected Lactobacillus (37.11%), Streptococcus (21.11%), and Staphylococcus (7.11%). Furthermore, our investigation revealed an abundance of auxiliary metabolic genes (AMGs) involved in carbon, nitrogen, and phosphorus cycling within the virome. We also detected a total of 113 antibiotic resistance genes (ARGs), covering major classes of antibiotics across all samples analyzed. Additionally, our findings indicated the presence of virulence factors including the clpP gene accounting for approximately 4.78%, along with toxin genes such as the RecT gene representing approximately 73.48% of all detected virulence factors and toxin genes among all samples analyzed. This study expands our understanding regarding both pathogenic bacteria and viruses present within sewage sludge while providing valuable insights into their ecological functions.


Subject(s)
Bacteria , Metagenomics , Sewage , Viruses , Wastewater , Wastewater/virology , Wastewater/microbiology , Bacteria/genetics , Bacteria/classification , Bacteria/isolation & purification , Sewage/virology , Sewage/microbiology , Humans , Viruses/genetics , Viruses/classification , Viruses/isolation & purification , Metagenome , Bacteriophages/genetics , Bacteriophages/isolation & purification , Bacteriophages/classification , Virome/genetics , Water Purification , Animals
8.
J Water Health ; 22(4): 717-720, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38678424

ABSTRACT

The measurement of the enterovirus and the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in sewage water is relevant in the early detection of the introduction or disappearance of these viruses in the ecosystem. We evaluated the co-circulation of the enteroviruses and SARS-CoV-2 in 81 sewage water samples collected between September 2021 and April 2023 from different regions of north and southeast Romania, at the border with Ukraine. We used, for the molecular detection of the pathogens, the multiplex real-time polymerase chain reaction (PCR) assay produced for respiratory samples and the Respiratory 2.1 Plus panel Biofire Film array. The isolation of enteroviruses was performed on cell culture lines, in accordance with the World Health Organization (WHO) recommendations. By molecular investigations, we detected the SARS-CoV-2 in 22 (27%) samples, and the human rhinovirus/enterovirus in 64 (79%) samples. By isolation on cell culture lines, 27 samples (33,33%) were positive for non-polio enteroviruses, and no poliovirus strains were isolated, proving the maintenance of the polio-free status in Romania. In an emergency situation, the molecular detection of the pathogens in sewage water using a PCR system integrating sample preparation, amplification, detection, and analysis in 1 h could be implemented.


Subject(s)
COVID-19 , Enterovirus , Poliomyelitis , SARS-CoV-2 , Sewage , Humans , Sewage/virology , Enterovirus/isolation & purification , Enterovirus/genetics , SARS-CoV-2/isolation & purification , Poliomyelitis/virology , Poliomyelitis/epidemiology , COVID-19/virology , COVID-19/epidemiology , Romania/epidemiology
9.
Sci Rep ; 14(1): 9251, 2024 04 22.
Article in English | MEDLINE | ID: mdl-38649443

ABSTRACT

The increasing prevalence of methicillin-resistant Staphylococcus aureus (MRSA) emphasises the urgent need for novel antimicrobial agents as alternatives to antibiotics. Bacteriophage therapy is one of the most promising antimicrobial strategies. Here, we isolated and comprehensively characterized a novel Staphylococcus phage, vB_SauM_VL10 (VL10), from urban sewage. The VL10 genome displays 141,746 bp of linear double-stranded DNA, containing 193 open reading frames and lacking tRNA, virulence, or antibiotic resistance genes. Phylogenetic analysis categorizes VL10 as a novel species within the Silviavirus genus, Twortvirinae subfamily. VL10 exhibits lytic behaviour characterized by efficient adsorption, a short latent period, and substantial burst size, with environmental stability. It demonstrates lytic activity against 79.06% of tested S. aureus strains, highlighting its species specificity. Additionally, VL10 effectively targets MRSA biofilms, reducing biomass and viable cells. In MRSA-infected G. mellonella larvae, VL10 enhances survival rates, supporting its potential for phage therapy applications. Moreover, the emergence of VL10-resistant S. aureus strains associated with fitness trade-offs, including reduced growth, biofilm formation, and virulence. Altogether, these findings emphasize VL10 as a promising candidate for developing therapeutic agents against MRSA infections, providing insights into phage biology and resistance dynamics.


Subject(s)
Biofilms , Genome, Viral , Methicillin-Resistant Staphylococcus aureus , Phylogeny , Staphylococcus Phages , Methicillin-Resistant Staphylococcus aureus/drug effects , Methicillin-Resistant Staphylococcus aureus/virology , Staphylococcus Phages/genetics , Biofilms/drug effects , Staphylococcal Infections/microbiology , Staphylococcal Infections/therapy , Staphylococcal Infections/drug therapy , Phage Therapy , Sewage/microbiology , Sewage/virology , Animals , Humans , Anti-Bacterial Agents/pharmacology
10.
J Virol Methods ; 327: 114918, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38556176

ABSTRACT

Wastewater-based epidemiology has allowed tracking the magnitude and distribution of SARS-CoV-2 in communities, allowing public health officials to prepare for impending outbreaks. While many factors influence recovery of SARS-CoV-2 from wastewater, proper extraction, concentration, and purification of RNA are key steps to ensure accurate detection of viral particles. The aim of this study was to compare the efficiency of four commonly used RNA extraction methods for detection of the SARS-CoV-2 RNA genome in sewage samples artificially inoculated with the virus, in order to identify a protocol that improves viral recovery. These methods included CTAB-based, TRIzol-based, and guanidinium thiocyanate (GTC)-based extraction procedures coupled with silica spin column-based purification, and an automated extraction/purification protocol using paramagnetic particles. Following RNA extraction, virus recovery rates were compared using RT-qPCR-based detection. The CTAB-based approach yielded the highest recovery rates and was the only method to consistently demonstrate stable virus recovery percentages regardless of the specific physicochemical characteristics of the samples tested. The TRIzol method proved to be the second most effective, yielding significantly higher recovery rates compared to both the GTC-based and the automated extraction methods. These results suggest that the CTAB-based approach could be a useful tool for the recovery of viral RNA from complex wastewater matrices.


Subject(s)
Cetrimonium , RNA, Viral , SARS-CoV-2 , Wastewater , Wastewater/virology , RNA, Viral/isolation & purification , RNA, Viral/genetics , SARS-CoV-2/isolation & purification , SARS-CoV-2/genetics , Cetrimonium/chemistry , Humans , Cetrimonium Compounds/chemistry , COVID-19/virology , COVID-19/diagnosis , Thiocyanates , Sewage/virology , Guanidines
11.
J Virol ; 97(10): e0082823, 2023 10 31.
Article in English | MEDLINE | ID: mdl-37747236

ABSTRACT

IMPORTANCE: Reoviruses infect many mammals and are widely studied as a model system for enteric viruses. However, most of our reovirus knowledge comes from laboratory strains maintained on immortalized L929 cells. Herein, we asked whether naturally circulating reoviruses possess the same genetic and phenotypic characteristics as laboratory strains. Naturally circulating reoviruses obtained from sewage were extremely diverse genetically. Moreover, sewage reoviruses exhibited poor fitness on L929 cells and relied heavily on gut proteases for viral uncoating and productive infection compared to laboratory strains. We then examined how naturally circulating reoviruses might adapt to cell culture conditions. Within three passages, virus isolates from the parental sewage population were selected, displaying improved fitness and intracellular uncoating in L929 cells. Remarkably, selected progeny clones were present at 0.01% of the parental population. Altogether, using reovirus as a model, our study demonstrates how the high genetic diversity of naturally circulating viruses results in rapid adaptation to new environments.


Subject(s)
Adaptation, Physiological , Genetic Fitness , Genome, Viral , Host Microbial Interactions , Peptide Hydrolases , Reoviridae , Virus Uncoating , Animals , Mice , Genome, Viral/genetics , Genomics , L Cells , Peptide Hydrolases/metabolism , Reoviridae/classification , Reoviridae/genetics , Reoviridae/metabolism , Serial Passage , Sewage/virology
12.
Environ Sci Pollut Res Int ; 30(10): 25050-25057, 2023 Feb.
Article in English | MEDLINE | ID: mdl-34138435

ABSTRACT

The viral RNA of SARS-Coronavirus-2 is known to be contaminating municipal wastewater. We aimed to assess if COVID-19 disease is spreading through wastewater. We studied the amount of viral RNA in raw sewage and the efficiency of the sewage treatment to remove the virus. Sewage water was collected before and after the activated sludge process three times during summer 2020 from three different sewage treatment plants. The sewage treatment was efficient in removing SARS-CoV-2 viral RNA. Each sewage treatment plant gathered wastewater from one hospital, of which COVID-19 admissions were used to describe the level of disease occurrence in the area. The presence of SARS-CoV-2 viral RNA-specific target genes (N1, N2, and E) was confirmed using RT-qPCR analysis. However, hospital admission did not correlate significantly with viral RNA. Moreover, viral RNA loads were relatively low, suggesting that sewage might preserve viral RNA in a hot climate only for a short time.


Subject(s)
COVID-19 , SARS-CoV-2 , Wastewater , Humans , COVID-19/epidemiology , RNA, Viral/isolation & purification , SARS-CoV-2/isolation & purification , Saudi Arabia/epidemiology , Sewage/virology , Wastewater/virology
13.
Viruses ; 14(12)2022 12 07.
Article in English | MEDLINE | ID: mdl-36560736

ABSTRACT

The genogroup II genotype 4 (GII.4) noroviruses are a major cause of viral gastroenteritis. Since the emergence of the Sydney_2012 variant, no novel norovirus GII.4 variants have been reported. The high diversity of noroviruses and periodic emergence of novel strains necessitates continuous global surveillance. The aim of this study was to assess the diversity of noroviruses in selected wastewater samples from Pretoria, South Africa (SA) using amplicon-based next-generation sequencing (NGS). Between June 2018 and August 2020, 200 raw sewage and final effluent samples were collected fortnightly from two wastewater treatment plants in Pretoria. Viruses were recovered using skimmed milk flocculation and glass wool adsorption-elution virus recovery methods and screened for noroviruses using a one-step real-time reverse-transcription PCR (RT-PCR). The norovirus BC genotyping region (570-579 bp) was amplified from detected norovirus strains and subjected to Illumina MiSeq NGS. Noroviruses were detected in 81% (162/200) of samples. The majority (89%, 89/100) of raw sewage samples were positive for at least one norovirus, compared with 73% (73/100) of final effluent samples. Overall, a total of 89 different GI and GII RdRp-capsid combinations were identified, including 51 putative novel recombinants, 34 previously reported RdRp-capsid combinations, one emerging novel recombinant and three Sanger-sequencing confirmed novel recombinants.


Subject(s)
Norovirus , Sewage , Wastewater , Humans , Caliciviridae Infections , Gastroenteritis/virology , Genotype , High-Throughput Nucleotide Sequencing , Molecular Epidemiology , Norovirus/genetics , Norovirus/isolation & purification , Phylogeny , RNA-Dependent RNA Polymerase/genetics , Sewage/virology , South Africa/epidemiology , Wastewater/virology , Reassortant Viruses/genetics , Reassortant Viruses/isolation & purification
15.
Sci Total Environ ; 851(Pt 2): 158350, 2022 Dec 10.
Article in English | MEDLINE | ID: mdl-36041621

ABSTRACT

Wastewater-based epidemiology (WBE) has been suggested as a useful tool to predict the emergence and investigate the extent of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In this study, we screened appropriate population biomarkers for wastewater SARS-CoV-2 normalization and compared the normalized SARS-CoV-2 values across locations with different demographic characteristics in southeastern Michigan. Wastewater samples were collected between December 2020 and October 2021 from nine neighborhood sewersheds in the Detroit Tri-County area. Using reverse transcriptase droplet digital polymerase chain reaction (RT-ddPCR), concentrations of N1 and N2 genes in the studied sites were quantified, with N1 values ranging from 1.92 × 102 genomic copies/L to 6.87 × 103 gc/L and N2 values ranging from 1.91 × 102 gc/L to 6.45 × 103 gc/L. The strongest correlations were observed with between cumulative COVID-19 cases per capita (referred as COVID-19 incidences thereafter), and SARS-CoV-2 concentrations normalized by total Kjeldahl nitrogen (TKN), creatinine, 5-hydroxyindoleacetic acid (5-HIAA) and xanthine when correlating the per capita SARS-CoV-2 and COVID-19 incidences. When SARS-CoV-2 concentrations in wastewater were normalized and compared with COVID-19 incidences, the differences between neighborhoods of varying demographics were reduced as compared to differences observed when comparing non-normalized SARS-CoV-2 with COVID-19 cases. This indicates when studying the disease burden in communities of different demographics, accurate per capita estimation is of great importance. The study suggests that monitoring selected water quality parameters or biomarkers, along with RNA concentrations in wastewater, will allow adequate data normalization for spatial comparisons, especially in areas where detailed sanitary sewage flows and contributing populations in the catchment areas are not available. This opens the possibility of using WBE to assess community infections in rural areas or the developing world where the contributing population of a sample could be unknown.


Subject(s)
COVID-19 , SARS-CoV-2 , Sewage , Humans , COVID-19/epidemiology , Creatinine , Hydroxyindoleacetic Acid , Incidence , Nitrogen , RNA , SARS-CoV-2/isolation & purification , Sewage/virology , United States , Wastewater , Xanthines
16.
Science ; 375(6585): 1100-1104, 2022 Mar 11.
Article in English | MEDLINE | ID: mdl-35271341
17.
Viruses ; 14(2)2022 01 27.
Article in English | MEDLINE | ID: mdl-35215857

ABSTRACT

Vibrio parahaemolyticus causes aquatic vibriosis. Its biofilm protects it from antibiotics; therefore, a new different method is needed to control V. parahaemolyticus for food safety. Phage therapy represents an alternative strategy to control biofilms. In this study, the lytic Vibrio phage vB_VpaP_FE11 (FE11) was isolated from the sewers of Guangzhou Huangsha Aquatic Market. Electron microscopy analysis revealed that FE11 has a typical podovirus morphology. Its optimal stability temperature and pH range were found to be 20-50 °C and 5-10 °C, respectively. It was completely inactivated following ultraviolet irradiation for 20 min. Its latent period is 10 min and burst size is 37 plaque forming units/cell. Its double-stranded DNA genome is 43,397 bp long, with a G + C content of 49.24% and 50 predicted protein-coding genes. As a lytic phage, FE11 not only prevented the formation of biofilms but also could destroy the formed biofilms effectively. Overall, phage vB_VpaP_FE11 is a potential biological control agent against V. parahaemolyticus and the biofilm it produces.


Subject(s)
Bacteriophages/physiology , Biofilms/growth & development , Podoviridae/physiology , Vibrio parahaemolyticus/physiology , Vibrio parahaemolyticus/virology , Bacteriolysis , Bacteriophages/classification , Bacteriophages/genetics , Bacteriophages/isolation & purification , Biological Control Agents , Genome, Viral , Host Specificity , Phage Therapy , Phylogeny , Podoviridae/classification , Podoviridae/genetics , Podoviridae/isolation & purification , Sewage/virology
18.
Viruses ; 14(2)2022 01 29.
Article in English | MEDLINE | ID: mdl-35215879

ABSTRACT

Salmonella and Escherichia coli (E. coli) food contamination could lead to serious foodborne diseases. The gradual increase in the incidence of foodborne disease invokes new and efficient methods to limit food pathogenic microorganism contamination. In this study, a polyvalent broad-spectrum Escherichia phage named Tequatrovirus EP01 was isolated from pig farm sewage. It could lyse both Salmonella Enteritidis (S. Enteritidis) and E. coli and exhibited broad host range. EP01 possessed a short latent period (10 min), a large burst size (80 PFU/cell), and moderate pH stability (4-10) and appropriate thermal tolerance (30-80 °C). Electron microscopy and genome sequence revealed that EP01 belonged to T4-like viruses genus, Myoviridae family. EP01 harbored 12 CDSs associated with receptor-binding proteins and lacked virulence genes and drug resistance genes. We tested the inhibitory effect of EP01 on S. Enteritidis, E. coli O157:H7, E. coli O114:K90 (B90), and E. coli O142:K86 (B) in liquid broth medium (LB). EP01 could significantly reduce the counts of all tested strains compared with phage-free groups. We further examined the effectiveness of EP01 in controlling bacterial contamination in two kinds of foods (meat and milk) contaminated with S. Enteritidis, E. coli O157:H7, E. coli O114:K90 (B90), and E. coli O142:K86 (B), respectively. EP01 significantly reduced the viable counts of all the tested bacteria (2.18-6.55 log10 CFU/sample, p < 0.05). A significant reduction of 6.55 log10 CFU/cm2 (p < 0.001) in bacterial counts on the surface of meat was observed with EP01 treatment. Addition of EP01 at MOI of 1 decreased the counts of bacteria by 4.3 log10 CFU/mL (p < 0.001) in milk. Generally, the inhibitory effect exhibited more stable at 4 °C than that at 28 °C, whereas the opposite results were observed in milk. The antibacterial effects were better at MOI of 1 than that at MOI of 0.001. These results suggests that phage EP01-based method is a promising strategy of controlling Salmonella and Escherichia coli pathogens to limit microbial food contamination.


Subject(s)
Escherichia coli/virology , Food Contamination/prevention & control , Myoviridae/physiology , Salmonella enteritidis/virology , Animals , Bacteriolysis , Escherichia coli/growth & development , Food Microbiology , Genome, Viral , Host Specificity , Meat/microbiology , Milk/microbiology , Myoviridae/classification , Myoviridae/genetics , Myoviridae/isolation & purification , Phylogeny , Salmonella enteritidis/growth & development , Sewage/virology , Swine
19.
Yakugaku Zasshi ; 142(1): 11-15, 2022.
Article in Japanese | MEDLINE | ID: mdl-34980746

ABSTRACT

The polio eradication program, launched in 1988, has successfully decreased the number of poliomyelitis patients worldwide. However, in areas with immunization gaps where oral polio vaccine coverage has dropped, outbreaks of more virulent vaccine-derived polioviruses (VDPVs) have become a threat to public health. In Japan, inactivated polio vaccine replaced oral polio vaccine as the routine immunization in 2012. Polio environmental surveillance (ES) has been conducted nationwide since 2013 to efficiently monitor the wild type poliovirus or VDPV, which may be imported from overseas. ES may also be utilized to detect other viruses in stool samples. We propose a method of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection based on the polio ES network, and establish a procedure to detect fragments of SARS-CoV-2 genome in wastewater solids. Our findings suggest that polio ES can be used to simultaneously monitor SARS-CoV-2 RNA fragments in sewage waters.


Subject(s)
Environmental Monitoring/methods , Poliovirus/isolation & purification , SARS-CoV-2/isolation & purification , Sewage/virology , Wastewater/virology , Disease Eradication , Humans , Japan , Poliovirus Vaccine, Inactivated , RNA, Viral/isolation & purification , SARS-CoV-2/genetics
20.
J Med Virol ; 94(4): 1315-1329, 2022 Apr.
Article in English | MEDLINE | ID: mdl-34825708

ABSTRACT

In December 2019, novel severe acute respiratory syndrome coronavirus 2 (nSARS-CoV-2) virus outbreaks emerged from Wuhan, China, and spread all over the world, including India. Molecular diagnosis of Coronavirus Disease 2019 (COVID) 19 for densely and highly populated countries like India is time-consuming. A few reports have described the successful diagnosis of nSARS-CoV-2 virus from sewage and wastewater samples contaminated with fecal matter, suggesting the diagnosis of COVID 19 from the same to raise an alarm about the community transmission of virus for implementation of evacuation and lockdown strategies. So far, the association between the detection of virus and its concentration in stool samples with severity of the disease and the presence or absence of gastrointestinal symptoms have been rarely reported. We led the search utilizing multiple databases, specifically PubMed (Medline), EMBASE, and Google Scholar. We conducted a literature survey on gastrointestinal infection and the spread of this virus through fecal-oral transmission. Reports suggested that the existence and persistence of nSARS-CoV-2 in anal/rectal swabs and stool specimens for a longer period of time than in nasopharyngeal swabs provides a strong tenable outcome of gastrointestinal contamination and dissemination of this infection via potential fecal-oral transmission. This review may be helpful to conduct further studies to address the enteric involvement and excretion of nSARS-CoV-2 RNA in feces and control the community spread in both COVID-19 patients ahead of the onset of symptoms and in asymptomatic individuals through wastewater and sewage surveillance as an early indication of infection. The existence of the viral genome and active viral particle actively participate in genomic variations. Hence, we comprehended the enteric spread of different viruses amongst communities with special reference to nSARS-CoV-2.


Subject(s)
COVID-19/virology , Disease Transmission, Infectious , Gastrointestinal Diseases/virology , SARS-CoV-2/isolation & purification , COVID-19/epidemiology , COVID-19/prevention & control , COVID-19/transmission , Disease Transmission, Infectious/prevention & control , Feces/virology , Gastrointestinal Diseases/epidemiology , Gastrointestinal Diseases/prevention & control , Gastrointestinal Tract/virology , Humans , India/epidemiology , SARS-CoV-2/genetics , Sewage/virology , Water Purification
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