scholarly journals Evaluating recovery, cost, and throughput of different concentration methods for SARS-CoV-2 wastewater-based epidemiology

2020 ◽  
Author(s):  
Zachary W. LaTurner ◽  
David M. Zong ◽  
Prashant Kalvapalle ◽  
Kiara Reyes Gamas ◽  
Austen Terwilliger ◽  
...  
Keyword(s):  
Mottle Virus ◽  
List Type ◽  
Pepper Mild Mottle Virus ◽  
Method Performance ◽  
Bovine Coronavirus ◽  
Bead Beating ◽  
Virus Rna ◽  
Recovery Cost ◽  
Wastewater Samples ◽  
Concentration Methods

AbstractAs the COVID-19 pandemic continues to affect communities across the globe, the need to contain the spread of the outbreaks is of paramount importance. Wastewater monitoring of the SARS-CoV-2 virus, the causative agent responsible for COVID-19, has emerged as a promising tool for health officials to anticipate outbreaks. As interest in wastewater monitoring continues to grow and municipalities begin to implement this approach, there is a need to further identify and evaluate methods used to concentrate SARS-CoV-2 virus RNA from wastewater samples. Here we evaluate the recovery, cost, and throughput of five different concentration methods for quantifying SARS-CoV-2 virus RNA in wastewater samples. We tested the five methods on six different wastewater samples. We also evaluated the use of a bovine coronavirus vaccine as a process control and pepper mild mottle virus as a normalization factor. Of the five methods we tested head-to-head, we found that HA filtration with bead beating performed the best in terms of sensitivity and cost. This evaluation can serve as a guide for laboratories establishing a protocol to perform wastewater monitoring of SARS-CoV-2.HighlightsFive methods for concentrating SARS-CoV-2 RNA from wastewater evaluatedMethod performance characterized via recovery, cost, throughput, and variabilityHA filtration with bead beating had highest recovery for comparatively low costBovine coronavirus, pepper mild mottle virus assessed as possible recovery controls

scholarly journals Pepper Mild Mottle Virus as an Indicator of Fecal Pollution

2009 ◽  
Vol 75 (22) ◽  
pp. 7261-7267 ◽  
Author(s):  
Karyna Rosario ◽  
Erin M. Symonds ◽  
Christopher Sinigalliano ◽  
Jill Stewart ◽  
Mya Breitbart
Keyword(s):  
The United States ◽  
Fecal Pollution ◽  
Point Sources ◽  
Mottle Virus ◽  
Treated Wastewater ◽  
Recreational Waters ◽  
Pepper Mild Mottle Virus ◽  
Fecal Samples ◽  
Seawater Samples ◽  
Wastewater Samples

ABSTRACT Accurate indicators of fecal pollution are needed in order to minimize public health risks associated with wastewater contamination in recreational waters. However, the bacterial indicators currently used for monitoring water quality do not correlate with the presence of pathogens. Here we demonstrate that the plant pathogen Pepper mild mottle virus (PMMoV) is widespread and abundant in wastewater from the United States, suggesting the utility of this virus as an indicator of human fecal pollution. Quantitative PCR was used to determine the abundance of PMMoV in raw sewage, treated wastewater, seawater exposed to wastewater, and fecal samples and/or intestinal homogenates from a wide variety of animals. PMMoV was present in all wastewater samples at concentrations greater than 1 million copies per milliliter of raw sewage. Despite the ubiquity of PMMoV in human feces, this virus was not detected in the majority of animal fecal samples tested, with the exception of chicken and seagull samples. PMMoV was detected in four out of six seawater samples collected near point sources of secondary treated wastewater off southeastern Florida, where it co-occurred with several other pathogens and indicators of fecal pollution. Since PMMoV was not found in nonpolluted seawater samples and could be detected in surface seawater for approximately 1 week after its initial introduction, the presence of PMMoV in the marine environment reflects a recent contamination event. Together, these data demonstrate that PMMoV is a promising new indicator of fecal pollution in coastal environments.

Pepper mild mottle virus in different water matrices

2020 ◽  
Vol 30 (Supplement_5) ◽  
Author(s):  
G Bonanno Ferraro ◽  
E Suffredini ◽  
P Mancini ◽  
C Veneri ◽  
M Iaconelli ◽  
...  
Keyword(s):  
Water Samples ◽  
Water Cycle ◽  
Human Subjects ◽  
Mottle Virus ◽  
Pepper Mild Mottle Virus ◽  
Healthy Human ◽  
Drinking Waters ◽  
Bathing Waters ◽  
Seawater Samples ◽  
Wastewater Samples

Abstract Background Pepper mild mottle virus (PMMoV), a plant virus belonging to Virgoviridae, has recently been suggested as a potential viral indicator for faecal pollution in aquatic environments, since it has been found to be abundantly excreted from healthy human subjects. Methods The occurrence, amount and diversity of PMMoV was investigated in water environments by nested RT-PCR and TaqMan based quantitative PCR. During 2017-2019, 251 water samples (92 urban wastewaters, 32 treated effluents, 16 surface water samples, 9 estuarine samples, 20 seawater samples, 67 groundwater samples, and 15 drinking waters) were analysed. Results PMMoV was detected in 73/92 (79%) wastewater samples, 22/32 (69%) treated sewages, 11/16 (69%) river samples, 6/9 (67%) estuarine samples, 5/20 (25%) bathing waters, and 9/67 (13%) groundwaters, whilst drinking water samples tested always negative. Mean viral concentrations (genome copies/L) were: raw sewage 4.2 × 106, treated sewage 7.4 × 105, river 3.2 × 103, estuarine waters 9.6 × 102, seawaters 3.0 × 102, groundwaters 7.7 × 101. Conclusions This study highlights the significant occurrence of PMMoV in aquatic environment in Italy, and a clear gradient of viral prevalence and concentrations from polluted to clean waters (wastewaters to drinking waters). Key messages PMMoV is ubiquitous throughout the water cycle in Italy with different concentrations. Studies are needed to evaluate the suitability of PMMoV as a viral indicator for human fecal pollution in waters.

Variability and evolution of the plant RNA virus pepper mild mottle virus.

1989 ◽  
Vol 63 (5) ◽  
pp. 2198-2203 ◽  
Author(s):  
E Rodríguez-Cerezo ◽  
A Moya ◽  
F García-Arenal
Keyword(s):  
Rna Virus ◽  
Mottle Virus ◽  
Pepper Mild Mottle Virus

scholarly journals The coat protein of tobamovirus acts as elicitor of both L 2 and L 4 gene-mediated resistance in Capsicum

2004 ◽  
Vol 85 (7) ◽  
pp. 2077-2085 ◽  
Author(s):  
P. Gilardi ◽  
I. García-Luque ◽  
M. T. Serra
Keyword(s):  
Hypersensitive Response ◽  
Host Response ◽  
Expression System ◽  
Potato Virus X ◽  
Mottle Virus ◽  
Coat Proteins ◽  
Pepper Mild Mottle Virus ◽  
Defence Genes ◽  
Glucuronidase Reporter ◽  
Capsicum Chacoense

In Capsicum, the resistance conferred by the L 2 gene is effective against all of the pepper-infecting tobamoviruses except Pepper mild mottle virus (PMMoV), whereas that conferred by the L 4 gene is effective against them all. These resistances are expressed by a hypersensitive response, manifested through the formation of necrotic local lesions (NLLs) at the primary site of infection. The Capsicum L 2 gene confers resistance to Paprika mild mottle virus (PaMMV), while the L 4 gene is effective against both PaMMV and PMMoV. The PaMMV and PMMoV coat proteins (CPs) were expressed in Capsicum frutescens (L 2 L 2) and Capsicum chacoense (L 4 L 4) plants using the heterologous Potato virus X (PVX)-based expression system. In C. frutescens (L 2 L 2) plants, the chimeric PVX virus containing the PaMMV CP was localized in the inoculated leaves and produced NLLs, whereas the chimeric PVX containing the PMMoV CP infected the plants systemically. Thus, the data indicated that the PaMMV CP is the only tobamovirus factor required for the induction of the host response mediated by the Capsicum L 2 resistance gene. In C. chacoense (L 4 L 4) plants, both chimeric viruses were localized to the inoculated leaves and produced NLLs, indicating that either PaMMV or PMMoV CPs are required to elicit the L 4 gene-mediated host response. In addition, transient expression of PaMMV CP into C. frutescens (L 2 L 2) leaves and PMMoV CP into C. chacoense (L 4 L 4) leaves by biolistic co-bombardment with a β-glucuronidase reporter gene led to the induction of cell death and the expression of host defence genes in both hosts. Thus, the tobamovirus CP is the elicitor of the Capsicum L 2 and L 4 gene-mediated hypersensitive response.

scholarly journals Elimination of pepper mild mottle virus from infected tobacco (Nicotiana benthamiana L.) plants by callus culture and the sieving technique

2012 ◽  
Vol 48 (6) ◽  
pp. 595-599 ◽  
Author(s):  
Youngsun Kwon ◽  
Md. Alamgir Kabir ◽  
Hong Wei Wang ◽  
Thirupathi Karuppanapandian ◽  
Jun-Cheol Moon ◽  
...  
Keyword(s):  
Callus Culture ◽  
Nicotiana Benthamiana ◽  
Mottle Virus ◽  
Pepper Mild Mottle Virus ◽  
Infected Tobacco

scholarly journals Pepper mild mottle virus as a water quality indicator

2018 ◽  
Vol 1 (1) ◽  
Author(s):  
Masaaki Kitajima ◽  
Hannah P. Sassi ◽  
Jason R. Torrey
Keyword(s):  
Water Quality ◽  
Quality Indicator ◽  
Mottle Virus ◽  
Pepper Mild Mottle Virus ◽  
Water Quality Indicator

scholarly journals Pepper Mild Mottle Virus Coat Protein Alone Can Elicit the Capsicum spp. L3 Gene-Mediated Resistance

1998 ◽  
Vol 11 (12) ◽  
pp. 1253-1257 ◽  
Author(s):  
P. Gilardi ◽  
I. García-Luque ◽  
M. T. Serra
Keyword(s):  
Coat Protein ◽  
Resistance Gene ◽  
Protein Gene ◽  
Expression System ◽  
Potato Virus X ◽  
Mottle Virus ◽  
Capsicum Chinense ◽  
Pepper Mild Mottle Virus ◽  
Capsicum Spp ◽  
Virus Coat

The pepper mild mottle virus (PMMoV-S) (an L3 hypersensitive response [HR]-inducer strain) coat protein was expressed in Capsicum chinense (L3L3) plants with the heterologous potato virus X (PVX)-based expression system. The chimeric virus was localized in the inoculated leaves and induced the HR, thus indicating that the tobamoviral sequences that affect induction of the HR conferred by the L3 resistance gene reside in the coat protein gene. Furthermore, transient expression of the PMMoV-S coat protein in C. chinense leaves by biolistic co-bombardment with a plasmid expressing the β-glucuronidase (GUS) gene leads to the induction of cell death and expression of host defense genes. Thus, the coat protein of PMMoV-S is the elicitor of the Capsicum spp. L3 resistance gene-mediated HR.

scholarly journals Detection and molecular characterization of Pepper mild mottle virus in Serbia

Genetika ◽  
2015 ◽  
Vol 47 (2) ◽  
pp. 651-663 ◽  
Author(s):  
Dragana Milosevic ◽  
Ivana Stankovic ◽  
Aleksandra Bulajic ◽  
Maja Ignjatov ◽  
Zorica Nikolic ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Molecular Characterization ◽  
Great Influence ◽  
Alfalfa Mosaic Virus ◽  
Resistance Breeding ◽  
Elisa Test ◽  
Mottle Virus ◽  
Pepper Mild Mottle Virus ◽  
Das Elisa

During 2009 and 2010, a survey was conducted in pepper crops to detect the possible presence of Pepper mild mottle virus (PMMoV) in Serbia. A total of 239 pepper samples from 39 crops at 26 localities were collected and analyzed for the presence of PMMoV, Cucumber mosaic virus (CMV), Potato virus Y (PVY), and Alfalfa mosaic virus (AMV), using DAS-ELISA test. Although it was detected in a small percentage, PMMoV could pose a threat to pepper production in Serbia due to its rapid seed-borne spread. Presence of PMMoV was confirmed by serological and biological detection, followed by conventional reverse transcription RT-PCR, using primers specific for the RNA-dependent RNA polymerase (RdRp) and the coat protein (CP) genes. Molecular identification confirmed that the Serbian isolates belong to PMMoV pathotypes P1,2 which do not break the resistance gene L3. Reconstructed phylogenetic tree confirmed the allocation of the Serbian isolates together with the majority of PMMoV isolates which belong to pathotypes P1,2. This study represents the first serological and molecular characterization of PMMoV infection of pepper in Serbia, and provides important data on the population structure. The obtained data could have great influence on pepper production in Serbia as well as future pepper resistance breeding in the country.

scholarly journals Enteric Viruses and Pepper Mild Mottle Virus Show Significant Correlation in Select Mid-Atlantic Agricultural Waters

2021 ◽  
Author(s):  
Brienna L. Anderson-Coughlin ◽  
Shani Craighead ◽  
Alyssa Kelly ◽  
Samantha Gartley ◽  
Adam Vanore ◽  
...  
Keyword(s):  
United States ◽  
Surface Waters ◽  
Irrigation Water ◽  
Water Samples ◽  
Enteric Viruses ◽  
The United States ◽  
Mottle Virus ◽  
Pepper Mild Mottle Virus ◽  
Atlantic Region ◽  
Viral Pathogen

Enteric viruses (EV) are the largest contributors to foodborne illness and outbreaks globally. Their ability to persist in the environment, coupled with the challenges experienced in environmental monitoring create a critical aperture through which agricultural crops may become contaminated. This study involved a seventeen-month investigation of select human enteric viruses and viral indicators in non-traditional irrigation water sources, surface and reclaimed waters, in the Mid-Atlantic region of the United States. Real-time quantitative PCR was used for detection of Aichi virus, hepatitis A virus, noroviruses GI and GII. Pepper mild mottle virus (PMMoV), a common viral indicator of human fecal contamination, was also evaluated along with atmospheric (air and water temperature, cloud cover, and precipitation 24 h, 7 d and 14 d prior to sample collection) and physicochemical (dissolved oxygen, pH, salinity and turbidity) data to determine if there were any association between EV and measured parameters. EV were detected more frequently in reclaimed waters (32%, n=22) than in surface waters (4%, n=49) similar to PMMoV detection frequency in surface (33%, n=42) and reclaimed (67%, n=21) water. Our data show a significant correlation between EV and PMMoV (R2=0.628; p<0.05) detection in reclaimed water samples, but not in surface waters (R2=0.476; p=0.78). Water salinity significantly affected the detection of both EV and PMMoV (p<0.05) as demonstrated by logistic regression analyses. These results provide relevant insights into the extent and degree of association between human enteric (pathogenic) viruses and water quality data in Mid-Atlantic surface and reclaimed waters as potential sources for agricultural irrigation. IMPORTANCE Microbiological analysis of agricultural waters is fundamental to ensure microbial food safety. The highly variable nature of non-traditional sources of irrigation water are particularly difficult to test for the presence of viruses. Multiple characteristics influence viral persistence in a water source as well as affect the recovery and detection methods which are employed. Testing for a suite of viruses in water samples is often too costly and labor intensive, making identification of suitable indicators for viral pathogen contamination necessary. The results from this study address two critical data gaps: enteric virus prevalence in surface and reclaimed waters of the Mid-Atlantic region of the United States and subsequent evaluation of physicochemical and atmospheric parameters used to inform the potential for use of indicators of viral contamination.

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