SARS-CoV-2 surveillance with environmental surface sampling in public areas.

Contaminated surfaces are one of the ways that coronavirus disease 2019 (COVID-19) may be transmitted. SARS-CoV-2 can be detected on environmental surfaces; however, few environmental sampling studies have been conducted in nonclinical settings. The objective of this study was to detect SARS-CoV-2 RNA on environmental surfaces in public areas in Las Vegas, Nevada. In total, 300 surface samples were collected from high-touch surfaces from high-congregate public locations and from a public health facility (PHF) that was visited by COVID-19 patients. Environmental samples were analyzed with quantitative reverse-transcriptase polymerase chain reaction (RT-qPCR) using SARS-CoV-2 specific primers and probes for three target genes. Results showed that 31 out of 300 (10.3%) surface samples tested positive for SARS-CoV-2, 24 at the PHF and 7 in high-congregate public locations. Concentrations ranged from 102 to 106 viral particles per 3 ml sample on a wide variety of materials. The data also showed that the N gene assay had greater sensitivity compared to the S and ORF gene assays. Besides frequently touched surfaces, SARS-CoV-2 was detected in restrooms, on floors and surfaces in contact with floors, as well as in a mop water sample. The results of this study describe the extent and distribution of environmental SARS-CoV-2 contamination in public areas in Las Vegas, Nevada. A method using the N gene PCR assay was developed for SARS-CoV-2 environmental monitoring in public areas. Environmental monitoring with this method can determine the specific sites of surface contamination in the community and may be beneficial for prevention of COVID-19 indirect transmission, and evaluation and improvement of infection control practices in public areas, public health facilities, universities, and businesses.

Evaluation of two surface sampling methods for the detection of Bacillus atrophaeus aerosolized in a test chamber.

In response to the airborne release of biothreat agents, surface sampling is often used to provide information on bioaerosol dispersal and deposition, to identify biocontaminant sources, and determine the effectiveness of decontamination. The objective of this project was to use aerosolization and deposition of dry spores to evaluate the efficiency of the cellulose sponge wipe and 37-mm cassette micro vacuum surface sampling methods for the collection of microorganisms from two contaminated surfaces, metal and concrete. Aerosolization trials were performed in a room-sized test chamber with known airborne concentrations of Bacillus atrophaeus spores serving as a surrogate for a bioterrorism agent. Following each aerosolization trial, the chamber heating, ventilation, air conditioning (HVAC) system was turned off to allow airborne spores to settle onto the test materials. Surface sampling was conducted and culture analysis was used to determine the concentration of B. atrophaeus on the surfaces. Results were compared with reference samples to determine the collection efficiency of the sampling methods. The sponge wipe sampling method was significantly more effective than the vacuum method for the collection of B. atrophaeus from both metal and concrete surfaces (P < 0.001). The collection efficiency of the sponge wipe method was 39.5% for metal and 26.5% for concrete, while the collection efficiency of the vacuum method was 7.6% for metal and 9.3% for concrete. The results of this study provided data on the collection efficiencies of two surface sampling methods for detection and enumeration of biocontaminants and can aid in selection of sampling methods.

Incidence of Neonatal Abstinence Syndrome Epidemic and Associated Predictors in Nevada: A Statewide Audit, 2016-2018.

Neonatal abstinence syndrome (NAS) is a postnatal withdrawal syndrome among neonates born to mothers with drug dependence disorders. NAS poses a significant public health challenge nationally, with a six-fold increase in incidence (1.2 to 6.7 per 1000 hospital births/year) from 2000-2016. Besides national data, it is critical to quantify NAS at the state-level to identify target areas for prevention. The objectives of this study were to ascertain statewide burden, including county and regional distribution of NAS in Nevada during 2016-2018, and to investigate potential factors associated with NAS. This study utilized hospital administrative data, and a total of 100,845 inpatient pediatric discharges were examined to identify NAS cases. Statistical analyses included estimation of crude incidence rates per 1000 hospital births and multilevel logistic regression modeling. NAS incidence in Nevada decreased slightly from 8.6 to 7.7 per 1000 hospital births between 2016 and 2018, but the overall incidence of 8 was substantially higher than earlier estimates (4.8/1000 hospital births) reported for 2013. Incidence was disproportionately higher among white newborns (12, 95% CI 11.0,13.0) and Medicaid enrollees (13.2, 95% CI 11.0,15.0). Southern Nevada had the highest incidence rate of 8.2 per 1000 hospital births. Nearly 75% of NAS cases were residents of Clark County. Incidence rates of NAS parallel the growing opioid prescription rates in Nevada and highlight the need for adopting opioid control prescribing practices to combat this drug epidemic. These findings might help in designing and evaluating state- and system-level interventions introduced to combat the opioid epidemic.

Behavioral Interventions to Increase Condom Use Among College Students in the United States: A Systematic Review.

Background. Traditional undergraduate college students in the United States are in the age range that experiences the highest rate of sexually transmitted infections (STIs) and are vulnerable to contracting STIs. Increasing condom use among college students is a prevention strategy to reduce the spread of STIs. Aim. The purpose of this systematic review of the literature was to identify behavioral interventions that increased condom use behaviors and/or intentions among college students. Method. The Preferred Reporting Items for Systematic Review and Meta-Analyses (PRISMA) guidelines were followed in systematically searching, extracting, appraising, and synthesizing the evidence. A quality assessment was also conducted with the tool provided by the Effective Public Health Practice Project. Results. The initial search yielded 715 records. After critical appraisal, seven articles remained for review. Discussion. Four of the interventions were developed using the three constructs of the information, motivation, and behavioral skills model, and all four found significant increases in condom use or condom use intentions. Additionally, interventions that included modules to increase self-efficacy for condom use, taught participants where to get condoms and how to negotiate condom use with partners, or elicited positive associations (feels) toward condoms saw increased condom use or intention to use condoms.

Comparison of Clinical & Laboratory Standards Institute standards in antimicrobial susceptibility among the carbapenemase producing Enterobacteriaceae.

AIM: Carbapenems are antibiotics reserved for treatment of severe infections. Carbapenem antimicrobial susceptibility testing profiles were determined in a population of Klebsiella pneumoniae, and their resistance assessed based on previous and current Clinical and Laboratory Standards Institute criteria. MATERIALS & METHODS: Isolates were examined using an automated antimicrobial susceptibility testing method, and real time polymerase chain reaction to detect the resistance (blaKPC) gene. RESULTS: The prevalence of blaKPC gene was 45/54 (83.3%). Five isolates that were susceptible under the previous criteria changed to nonsusceptible with the current standards. The overall difference in susceptibility between the standards was 8%. CONCLUSION: This study shows that the current Clinical and Laboratory Standards Institute criteria may not offer additional benefits in the fight against carbapenem-resistant Enterobacteriaceae.

Interventions to Reduce Loss to Follow-up During All Stages of the HIV Care Continuum in Sub-Saharan Africa: A Systematic Review.

The continuum of care for successful HIV treatment includes HIV testing, linkage, engagement in care, and retention on antiretroviral therapy (ART). Loss to follow-up (LTFU) is a significant disruption to this pathway and a common outcome in sub-Saharan Africa. This review of literature identified interventions that have reduced LTFU in the HIV care continuum. A search was conducted utilizing terms that combined the disease state, stages of the HIV care continuum, interventions, and LTFU in sub-Saharan Africa and articles published between January 2010 and July 2015. Thirteen articles were included in the final review. Use of point of care CD4 testing and community-supported programs improved linkage, engagement, and retention in care. There are few interventions directed at LTFU and none that span across the entire continuum of HIV care. Further research could focus on devising programs that include a series of interventions that will be effective through the entire continuum.

Development of a polymerase chain reaction assay for the rapid detection of the oral pathogenic bacterium, Selenomonas noxia.

BACKGROUND: In recent studies, periodontal health has been linked to being overweight and/or obese. Among common oral bacteria, Selenomonas noxia has been implicated in converting periodontal health to disease, and Selenomonas species have also been found in gastric ulcers. The objective of this study was to develop and validate a quantitative polymerase chain reaction (qPCR) assay for the specific and rapid detection of S. noxia. METHODS: Two oligonucleotide primer pairs and one probe were designed and tested to determine optimal amplification signal with three strains of S. noxia. The PCR assay was tested against fourteen non-target organisms, including closely related oral Selenomonads, one phylogenetically closely related bacterium, and two commonly isolated oral bacteria. RESULTS: One of the primer sets was more sensitive at detecting the target organism and was selected for optimization and validation experiments. The designed primers and probe amplified the target organism with 100% specificity. PCR inhibition was observed with an internal positive control, and inhibition was resolved by diluting the DNA extract. CONCLUSIONS: The qPCR assay designed in this study can be used to specifically detect S. noxia in the clinical setting and in future research involving the enhanced detection of S. noxia. The assay can also be used in epidemiological studies for understanding the role of S. noxia in disease processes including, but not limited to, oral health and obesity of infectious origin.

Development and evaluation of a real-time quantitative PCR assay for Aspergillus flavus.

Aspergillus flavus is a ubiquitous mold and the most common mold contaminating foodstuffs. Many strains of A. flavus produce aflatoxins. In addition it is an allergen and an opportunistic pathogen of animals and plants. A. flavus often is underestimated in traditional culture analyses due to the expertise required and the cost associated with speciating members of the genus Aspergillus. The goal of this study was to develop and validate a primer and probe set for the rapid detection and quantitation of A. flavus in pure culture using real-time quantitative polymerase chain reaction (QPCR) amplification. Unique DNA regions were located in the genome of the target organism by sequence comparison with the GenBank database, and several candidate oligonucleotides were identified from the scientific literature for potential use with the TaqMan QPCR technology. Three primer and probe sets were designed and validated for specificity and sensitivity in laboratory experiments. Initial screening to test for sensitivity was performed with seven A. flavus isolates and selected nontarget fungi. Specificity testing was conducted with the selected primer and probe set, which amplified all nine A. flavus isolates tested, including an aflatoxin producing strain. The primers did not amplify DNA extracted from 39 other fungal species (comprising 16 genera), including 18 other Aspergillus species and six Penicillium species. No amplification of human or bacterial DNA was observed; however cross-reactivity was observed with Aspergillus oryzae. PCR analysis of DNA dilutions and the use of an internal positive control demonstrated that 67% of the fungal DNA samples assayed contained PCR inhibitors. The assay validated for the target organism is capable of producing PCR results in less than 1 h after DNA extraction. The results of this research demonstrate the capabilities of QPCR for the enhanced detection and enumeration of fungi of significance to human health.

Evaluation of two surface sampling methods for detection of Erwinia herbicola on a variety of materials by culture and quantitative PCR.

This research was designed to evaluate surface sampling protocols for use with culture and quantitative PCR (QPCR) amplification assay for detection of the gram-negative bacterial biothreat simulant Erwinia herbicola on a variety of surface materials. Surfaces selected for evaluation were wood laminate, glass and computer monitor screens, metal file cabinets, plastic arena seats, nylon seat cushions, finished concrete flooring, and vinyl tile flooring. Laboratory and test chamber studies were performed to evaluate two sampling methods, a sponge and a macrofoam swab, for detection of E. herbicola on surface materials. In laboratory trials, seven materials were inoculated with a known concentration of E. herbicola cells and samples were collected from the surfaces of the materials to determine sampling efficiencies. Culture analysis was ineffective for assessing E. herbicola collection efficiency because very few culturable cells were obtained from surface samples. QPCR demonstrated that E. herbicola DNA was present in high concentrations on all of the surface samples, and sampling efficiencies ranged from 0.7 to 52.2%, depending on the sampling method and the surface material. The swab was generally more efficient than the sponge for collection of E. herbicola from surfaces. Test chamber trials were also performed in which E. herbicola was aerosolized into the chamber and allowed to settle onto test materials. Surface sampling results supported those obtained in laboratory trials. The results of this study demonstrate the capabilities of QPCR to enhance the detection and enumeration of biocontaminants on surface materials and provide information on the comparability of sampling methods.

Evaluation of the Biological Sampling Kit (BiSKit) for large-area surface sampling.

Current surface sampling methods for microbial contaminants are designed to sample small areas and utilize culture analysis. The total number of microbes recovered is low because a small area is sampled, making detection of a potential pathogen more difficult. Furthermore, sampling of small areas requires a greater number of samples to be collected, which delays the reporting of results, taxes laboratory resources and staffing, and increases analysis costs. A new biological surface sampling method, the Biological Sampling Kit (BiSKit), designed to sample large areas and to be compatible with testing with a variety of technologies, including PCR and immunoassay, was evaluated and compared to other surface sampling strategies. In experimental room trials, wood laminate and metal surfaces were contaminated by aerosolization of Bacillus atrophaeus spores, a simulant for Bacillus anthracis, into the room, followed by settling of the spores onto the test surfaces. The surfaces were sampled with the BiSKit, a cotton-based swab, and a foam-based swab. Samples were analyzed by culturing, quantitative PCR, and immunological assays. The results showed that the large surface area (1 m2) sampled with the BiSKit resulted in concentrations of B. atrophaeus in samples that were up to 10-fold higher than the concentrations obtained with the other methods tested. A comparison of wet and dry sampling with the BiSKit indicated that dry sampling was more efficient (efficiency, 18.4%) than wet sampling (efficiency, 11.3%). The sensitivities of detection of B. atrophaeus on metal surfaces were 42 +/- 5.8 CFU/m2 for wet sampling and 100.5 +/- 10.2 CFU/m2 for dry sampling. These results demonstrate that the use of a sampling device capable of sampling larger areas results in higher sensitivity than that obtained with currently available methods and has the advantage of sampling larger areas, thus requiring collection of fewer samples per site.

Determination of the efficacy of two building decontamination strategies by surface sampling with culture and quantitative PCR analysis.

The efficacy of currently available decontamination strategies for the treatment of indoor furnishings contaminated with bioterrorism agents is poorly understood. Efficacy testing of decontamination products in a controlled environment is needed to ensure that effective methods are used to decontaminate domestic and workplace settings. An experimental room supplied with materials used in office furnishings (i.e., wood laminate, painted metal, and vinyl tile) was used with controlled dry aerosol releases of endospores of Bacillus atrophaeus ("Bacillus subtilis subsp. niger," also referred to as BG), a Bacillus anthracis surrogate. Studies were performed using two test products, a foam decontaminant and chlorine dioxide gas. Surface samples were collected pre- and posttreatment with three sampling methods and analyzed by culture and quantitative PCR (QPCR). Additional aerosol releases with environmental background present on the surface materials were also conducted to determine if there was any interference with decontamination or sample analysis. Culture results indicated that 10(5) to 10(6) CFU per sample were present on surfaces before decontamination. After decontamination with the foam, no culturable B. atrophaeus spores were detected. After decontamination with chlorine dioxide gas, no culturable B. atrophaeus was detected in 24 of 27 samples (89%). However, QPCR analysis showed that B. atrophaeus DNA was still present after decontamination with both methods. Environmental background material had no apparent effect on decontamination, but inhibition of the QPCR assay was observed. These results demonstrate the effectiveness of two decontamination methods and illustrate the utility of surface sampling and QPCR analysis for the evaluation of decontamination strategies.

Detection and enumeration of airborne biocontaminants.

The sampling and analysis of airborne microorganisms has received attention in recent years owing to concerns with mold contamination in indoor environments and the threat of bioterrorism. Traditionally, the detection and enumeration of airborne microorganisms has been conducted using light microscopy and/or culture-based methods; however, these analyses are time-consuming, laborious, subjective and lack sensitivity and specificity. The use of molecular methods, such as quantitative polymerase chain reaction amplification, can enhance monitoring strategies by increasing sensitivity and specificity, while decreasing the time required for analysis.