Wastewater Knows Pathogen Spread: Analysis of Residential Wastewater for Infectious Microorganisms including SARS-CoV-2.

BACKGROUND: We aimed to identify the genes of 35 pathogens, including viruses, bacteria, and protozoa that cause waterborne infectious diseases, and to assess the feasibility of a wastewater-based surveillance system. MATERIALS AND METHODS: Wastewater was aseptically sampled once a month from 2 sites. A total of 1 L of wastewater from each site underwent 0.2 µm filtration to generate the sample A. Subsequently, 200 ul of the filtered water was ultra-filtered and concentrated to generate the sample B, which was mixed with sample A in a 1:1 ratio. We performed a Filmarray® Gastrointestinal (GI) panel (BioFire Diagnnostics', Salt Lake City, UT, USA) test to simultaneously detect 13 enterobacteria, 5 enterovirus, and 4 protozoa. RNA was extracted to assess 18 types of viruses. RESULTS: Severe acute respiratory syndrome coronavirus 2 adenovirus, bocavirus, and rhinovirus was detected at both site. Norovirus GI/GII was continuously detected at both sites. Moreover, adenovirus, group A rotaviruses, and hepatitis A virus were frequently detected; however, hepatitis E virus was absent at either site. Campylobacter, enteroaggregative Escherichia coli, enterotoxigenic E. coli, Shiga toxin-producing E. coli, and Clostridioides difficile toxin A/B were detected at both sites. Giardia lamblia was also detected in both sites. CONCLUSION: We analyze multiple infectious disease pathogens under sample surveillance with incidence. Further in-depth studies on wastewater-based surveillance will be feasible and important.

Effects of Non-Pharmacological Interventions on Respiratory Viruses Other Than SARS-CoV-2: Analysis of Laboratory Surveillance and Literature Review From 2018 to 2021.

BACKGROUND: Since the global coronavirus disease 2019 (COVID-19) pandemic, non-pharmacological interventions (NPIs) such as extensive and comprehensive hand hygiene, mask-wearing, and social distancing have been implemented globally. This study aimed to investigate changes in respiratory viruses other than severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) that occurred following the implementation of these NPIs. METHODS: From January 2018 to December 2021, influenza-like illness patient specimens and specimens from the Korea Influenza and Respiratory Viruses Surveillance System were analyzed at the Incheon Metropolitan City Institute of Public Health and Environment. Oropharyngeal or nasopharyngeal swab samples from respiratory infection patients were transferred in a virus transport medium at 4°C. After RNA or DNA extraction, respiratory virus-specific genes for human influenza virus (IFV), adenovirus (ADV), parainfluenza virus (PIV), respiratory syncytial virus (RSV), human rhinovirus (hRV), human coronavirus, human bocavirus, and human metapneumovirus were detected by individual real-time reverse transcription polymerase chain reaction. RESULTS: A total 3,334 samples were collected. After NPI was implemented, the detection of respiratory viruses other than SARS-CoV-2 decreased overall. The yearly detection rate of respiratory viruses was decreased from 69.5% (399/574) in 2018 and 73.3% (505/689) in 2019 to 19.8% (206/1,043) in 2020 and 34.9% (365/1,028) in 2021. The epidemic was more prominent in respiratory viruses such as IFV and RSV, which were considered dominant viruses, especially those with viral envelopes. Among viruses that were not considered dominant, hRV showed no clear change before and after NPI, while PIV showed a rapid increase compared to the existing dominant viruses between October-December 2021, after the increase in the number of gatherings started at the end of September and the "Relaxing COVID19 and mitigation policy," which was implemented on November 1. CONCLUSION: NPI seems to have influenced the isolation and transmission of respiratory viruses in South Korea. In the future, additional studies focusing on the isolation and transmission patterns of respiratory viruses following NPI are needed.

Investigation of Pathogenic Escherichia coli Contamination of Vegetables Distributed in a Korean Agricultural Wholesale Market.

The purpose of this study was to investigate the monthly contamination rate of pathogenic Escherichia coli, a major cause of food poisoning, in vegetables sold in agricultural wholesale markets, which distribute vegetables from all over the country, in the Incheon Metropolitan City area, South Korea, and to identify a source of the pathogen. In total, 1739 vegetables of 80 types, along with 109 soil, 67 manure, and 33 livestock feces samples, were tested for pathogenic E. coli using polymerase chain reaction, from September 2016 through August 2017. The average annual prevalence rate of vegetables was 5.8%, and the prevalence rate was above 5% from June through October. The highest prevalence rate (15.7%) was recorded in July. Water dropwort showed the highest prevalence rate (28.6%) among the vegetables examined. Pathogenic E. coli was detected in >20 types of the vegetables that were to be consumed without cooking. Among these, the prevalence rates of ponytail radish (n = 21), crown daisy (n = 86), young radish (n = 68), romaine lettuce (n = 133), perilla leaf (n = 103), Korean leek (n = 43), young Chinese cabbage (n = 68), and Chinese cabbage (n = 30) were 9.5%, 8.1%, 7.4%, 6.8%, 4.9%, 4.7%, 4.4%, and 3.3%, respectively. Among the vegetables cooked before consumption, prevalence rates were 28.6%, 27.3%, and 25.0% in wormwood, sweet potato stalk, and edible mountain vegetables (Saussurea sp., etc.), respectively. In soil, manure, and livestock feces, 36.7%, 26.9%, and 90.6% prevalence rates were confirmed, respectively. This study confirmed the pathogenic E. coli contamination of vegetables to be consumed without cooking. Therefore, to produce agricultural products that do not induce food poisoning and are safe for consumption, it is important to develop a process for killing the pathogenic microorganisms and set up a sanitary environment for effectively managing compost. In addition, it is necessary to establish surveillance systems to monitor the production chain.

Preliminary stable isotope analyses for propellant discrimination in shotshells.

RATIONALE: This study aimed to develop methods to determine the identity and trace the origin of propellants used in shotshells. Specifically, the use of organic component and stable isotope analysis techniques, such as bulk stable isotope analysis (BSIA) and compound-specific isotope analysis (CSIA) techniques, for the study of shotshell propellants was investigated. METHODS: Nine samples of shotshell propellants from different manufacturing countries and brands were analyzed for explosive and additive components by gas chromatography/mass spectrometry and thin-layer chromatography. BSIA of the propellants was achieved using elemental analysis/isotope ratio mass spectrometry without a pretreatment process. For the CSIA of nitroglycerin, double-base powder propellants were extracted with ether, and the isotope ratios of carbon and nitrogen were measured by gas chromatography/isotope ratio mass spectrometry. RESULTS: Nine samples drawn from seven brands in four countries were classified into five groups by organic component analysis, while eight classification groups were identified by BSIA. Thus, two samples could not be distinguished from each other by either BSIA or organic component analysis. Subsequently, with the use of results obtained with CSIA for nitroglycerin, all the samples could be classified into different groups. These findings suggest that the nine propellant samples were all composed of different ingredients or raw materials from different sources. CONCLUSIONS: Stable isotope ratio analyses were performed for propellant discrimination. The combined BSIA, CSIA and organic component analysis techniques were able to successfully distinguish the nine shotshell propellants from seven brands sourced from four different countries, and the results suggested that the samples contained different ingredients or raw materials from different sources. We therefore can conclude that reliable results can be obtained using combined isotope analysis methods such as CSIA and BSIA for origin tracing and identity determination.

Analysis of ethyl glucuronide and ethyl sulfate in blood to determine the absorption or elimination phase of alcohol for Korean.

This study was conducted to understand alcohol kinetics for Koreans and to determine whether an individual is in absorption phase or elimination phase at the time of blood collection by analyzing of ethyl glucuronide and ethyl sulfate in blood. A total of 50 healthy adults was selected and assigned to drink 1g of ethanol per kg body weight of individual within 1h. Blood samples were then collected every 15min for the first 3h, 30min next 3h, and 1h last 9h. Urine samples were also collected from the individual, but not under the controlled environment. All samples were then analyzed by gas chromatography with a flame ionization detector (GC-FID) for alcohol and liquid chromatography-mass/mass spectrometry (LC-MS/MS) for EtG and EtS. The maximum BAC (Cmax) was 0.138% (g/100mL) in average under the controlled experimental condition. Alcohol elimination rates (ß) in average were 0.020% for male and 0.024% for female, respectively. It was found that the ratio of UAC and BAC was less than 1 in the absorption phase and the average ratio of UAC and BAC was 1.47 in the elimination phase. The comparison of BAC (g/L) and EtG (mg/L) absorption and elimination curves showed that the intersection time was 3.9h in average. It is shown that the ratio of EtG (mg/L)/BAC (g/L) is higher than 1, the individual would be in elimination phase of BAC. At the time of Cmax, the ratio of EtG (mg/L)/BAC (g/L) was 0.255±0.132 (SD) in average.

Tracing the source of methomyl using stable isotope analysis.

RATIONALE: Pesticide self-poisoning is a major method of suicide in many agricultural communities worldwide. In addition, there are a number of known crime cases related to people being harmed by insecticides. Methomyl, a prohibited insecticide in the Republic of Korea, has high toxicity and is frequently used for self-poisoning. In this study, we investigated the source of origin of methamyl in a fatal poisoning case using stable isotope ratio analysis. METHODS: Two bottles of Soju from a crime scene were seized and nine different brands of methomyl products were collected from the neighborhood for analysis. In addition, the gastric contents and energy drink from the person who had committed suicide were analyzed. Isotope analysis using GC/Isolink/IRMS was conducted to determine the source of the methomyl by comparing their carbon and nitrogen isotope ratios. Linear discriminant analysis was utilized to verify the results. RESULTS: Isotope ratio analysis showed that the isotope ratio ranges of methomyl found in the Soju, the gastric contents of the suicide victim, and the energy drink bottle were similar to those of a seized methomyl product, Samgong methomyl 2011 (SG11). Thus, it was assumed that SG11 was used in this fatal poisoning case. CONCLUSIONS: This study demonstrates the potential of stable isotope ratio analysis for the determination of insecticide origin in fatal poisoning cases.

Classification and individualization of used engine oils using elemental composition and discriminant analysis.

The six most common commercial automotive gasoline and diesel engine oils in the Republic of Korea, ZIC A, ZIC XQ RV/SUV, Kixx G1, Kixx RV, and the brand name products HD Premium gasoline and HD Premium diesel, were randomly used in nineteen different vehicles. Samples of seventy-six used engine oils, which were withdrawn from the sumps of those vehicles at different intervals, were analyzed using inductively coupled plasma-optical emission spectrometry (ICP-OES), and statistically compared. Two data analysis strategies were used to interpret and understand the elemental profiles in the multi-dimensional data. Macro (additive elements of Ca, Zn and P) and trace (wear metal elements of Ag, Al, Ba, Cd, Cr, Cu, Fe, Mg, Mo, Na, Ni, Pb and Sn) elements were used as potential markers to determine the brand of oil used and the engine type in which the oil was used, and to trace the individual vehicle for forensic purposes. The discriminant analysis statistical technique was applied, and its prediction ability was assessed. In this study, 92.1%, 82.9% and 92.1% of the cross-validated grouped cases correctly predicted the brand of oil, the engine type and the vehicle that was the source of the oil, respectively.

Antioxidant and antigenotoxic activities of Korean fermented soybean.

This study was aimed at evaluating the antioxidative and antigenotoxic activities of Korean fermented soybean (Chungkookjang) in vitro and in vivo. The 100% ethanol extract of Chungkookjang (CKJ) inhibited the generation of 1,1-diphenyl-2-picryl hydrazine (DPPH) radicals, and had an inhibitory effect on LDL oxidation. CKJ and its constituents (genistein and daidzein) also inhibited H(2)O(2)-induced DNA damage from NIH/3T3 fibroblasts. Furthermore, they showed the cytoprotective effects against H(2)O(2)-induced cell death. In vivo study also demonstrated that an oral administration of CKJ extract (800 mg/kg/day) for 2 weeks potently inhibited the formation of malondialdehyde, the damage of DNA and the formation of micronucleated reticulocytes in KBrO(3)-treated mice. The well-known antioxidants, trolox and vitamin C, also showed the potent inhibition on these parameters. All these results indicate that CKJ extract may be a useful antigenotoxic antioxidant by scavenging free radicals, inhibiting lipid peroxidation and protecting against oxidative DNA damage. The isoflavones, genistein and daidzein, may contribute to these biological effects of CKJ extract at least in part. Korean fermented soybean (Chungkookjang) is suggested to be a promising functional food witch can prevent oxidative stress.