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Background We determined the burden of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in air and on surfaces in rooms of patients hospitalized with coronavirus disease 2019 (COVID-19) and investigated patient characteristics associated with SARS-CoV-2 environmental contamination. Methods Nasopharyngeal swabs, surface, and air samples were collected from the rooms of 78 inpatients with COVID-19 at 6 acute care hospitals in Toronto from March to May 2020. Samples were tested for SARS-CoV-2 ribonucleic acid (RNA), cultured to determine potential infectivity, and whole viral genomes were sequenced. Association between patient factors and detection of SARS-CoV-2 RNA in surface samples were investigated. Results Severe acute respiratory syndrome coronavirus 2 RNA was detected from surfaces (125 of 474 samples; 42 of 78 patients) and air (3 of 146 samples; 3 of 45 patients); 17% (6 of 36) of surface samples from 3 patients yielded viable virus. Viral sequences from nasopharyngeal and surface samples clustered by patient. Multivariable analysis indicated hypoxia at admission, polymerase chain reaction-positive nasopharyngeal swab (cycle threshold of ≤30) on or after surface sampling date, higher Charlson comorbidity score, and shorter time from onset of illness to sampling date were significantly associated with detection of SARS-CoV-2 RNA in surface samples. Conclusions The infrequent recovery of infectious SARS-CoV-2 virus from the environment suggests that the risk to healthcare workers from air and near-patient surfaces in acute care hospital wards is likely limited.

Smoking is considered as one of the major reasons behind genetic variations in cardiometabolic disorders. However, effect of nicotine via smoking on Pakistani population still needs to be elucidated. This study was aimed to investigate genetic variation among PPAR-γ, FTO, and ABCC8 genes in cardiometabolic patients along with their biochemical parameters. A total of 472 CVD patients were enrolled in this study and divided into three groups; n = 144 for PPAR-γ (C/G) variation and n = 164 in each group to investigate FTO (T/A) and ABCC8 (G/T) variation, respectively. Polymorphisms within groups were identified by using Tetra and/or Tri ARM-PCR. This study showed positive association among genetic polymorphisms in PPAR-γ, FTO, and ABCC8 groups with altered metabolic parameters in CVD patients. Findings showed that smoking is major contributory factor for genetic polymorphism that was strongly associated with elevated blood glucose and serum TGs accompanying PPAR-γ, FTO, and ABCC8 genetic polymorphism in 25%, 24%, and 20% in smokers and 11%, 10%, and 5% in non-smoker CVD patients, respectively. However, highest polymorphism occurred in PPAR-γ both in smokers and non-smoker CVD patients that show that smoking-mediated gene polymorphism might be a contributory factor in provoking CVD risk approximately twice in smokers as compared to that in non-smoker CVD patients.

To compare sensitivity of specimens for COVID-19 diagnosis, we tested 151 nasopharyngeal/midturbinate swab pairs from 117 COVID-19 inpatients using reverse-transcriptase polymerase chain reaction (RT-PCR). Sensitivity was 94% for nasopharyngeal and 75% for midturbinate swabs (P = .0001). In 88 nasopharyngeal/midturbinate pairs with matched saliva, sensitivity was 86% for nasopharyngeal swabs and 88% for combined midturbinate swabs/saliva.

SummaryBackgroundThe aim of this prospective cohort study was to determine the burden of SARS-CoV-2 in air and on surfaces in rooms of patients hospitalized with COVID-19, and to identify patient characteristics associated with SARS-CoV-2 environmental contamination.MethodsNasopharyngeal swabs, surface, and air samples were collected from the rooms of 78 inpatients with COVID-19 at six acute care hospitals in Toronto from March to May 2020. Samples were tested for SARS-CoV-2 viral RNA and cultured to determine potential infectivity. Whole viral genomes were sequenced from nasopharyngeal and surface samples. Association between patient factors and detection of SARS-CoV-2 RNA in surface samples were investigated using a mixed-effects logistic regression model.FindingsSARS-CoV-2 RNA was detected from surfaces (125/474 samples; 42/78 patients) and air (3/146 samples; 3/45 patients) in COVID-19 patient rooms; 17% (6/36) of surface samples from three patients yielded viable virus. Viral sequences from nasopharyngeal and surface samples clustered by patient.Multivariable analysis indicated hypoxia at admission, a PCR-positive nasopharyngeal swab with a cycle threshold of ≤30 on or after surface sampling date, higher Charlson co-morbidity score, and shorter time from onset of illness to sample date were significantly associated with detection of SARS-CoV-2 RNA in surface samples.InterpretationThe infrequent recovery of infectious SARS-CoV-2 virus from the environment suggests that the risk to healthcare workers from air and near-patient surfaces in acute care hospital wards is likely limited. Surface contamination was greater when patients were earlier in their course of illness and in those with hypoxia, multiple co-morbidities, and higher SARS-CoV-2 RNA concentration in NP swabs. Our results suggest that air and surfaces may pose limited risk a few days after admission to acute care hospitals.

Background Point-of-care tests (POCT) are promising tools to detect SARS-CoV-2 in specific settings. Initial reports suggest the ID NOW™ COVID-19 assay (Abbott Diagnostics Inc, USA) is less sensitive than standard real-time reverse transcription polymerase chain reaction (rRT-PCR) assays. This has raised concern over false negatives in SARS-CoV-2 POCT. Objectives We compared the performance of the ID NOW™ COVID-19 assay to our in-house rRT-PCR assay to assess whether dry swabs used in ID NOW™ testing could be stored in transport media and be re-tested by rRT-PCR for redundancy and to provide material for further investigation. Methods Paired respiratory swabs collected from patients at three acute care hospitals were used. One swab in transport media (McMaster Molecular Media (MMM)) was tested for SARS-CoV-2 by a laboratory-developed two-target rRT-PCR assay. The second was stored dry in a sterile container and tested by the ID NOW™ COVID-19 assay. Following ID NOW™ testing, dry swabs were stored in MMM for up to 48 h and re-tested by rRT-PCR. Serially diluted SARS-CoV-2 particles were used to assess the impact of heat inactivation and storage time. Results Respiratory swabs (n = 343) from 179 individuals were included. Using rRT-PCR results as the comparator, the ID NOW™ COVID-19 assay had positive (PPA) and negative (NPA) percent agreements of 87.0% (95% CI:0.74–0.94) and 99.7% (95% CI:0.98–0.99). Re-tested swabs placed in MMM following ID NOW testing had PPA and NPA of 88.8% (95% CI:0.76–0.95) and 99.7% (95% CI:0.98–0.99), respectively. Conclusions Storing spent dry swabs in transport media for redundancy rRT-PCR testing is a potential approach to address possible false negatives with the ID NOW™ COVID-19 assay.

Tobacco smoking is a leading concern to a global health; kills almost six million individuals annually and associated with life-threating health issues. Nearly 80% smokers live in middle- and low-income countries where illness and mortality rate due to cigarette smoking are higher. Tobacco smoking is interlinked with a numerous disease, comprising lungs cancer, chronic obstructive pulmonary disease, infertility, cardiovascular events, stroke, and metabolic disorders. Tobacco smoking has ability to obstruct and/or interfere in the function of endocrine system; has been entitled as endocrine disrupting chemicals (EDCs). EDCs are a heterogeneous group of exogenous compounds that can restrict with several facets of endogenous hormones. Standard role of endocrine system is reliant on hormonal/enzymatic pathway which may act as chemical and biological messengers to regulate the physiological functions of an organism. Tobacco smoking is also responsible to impair the regular metabolic pathway through interaction with members of superfamily of nuclear receptor such as peroxisome proliferator activated receptor, thyroid hormone receptors, liver X receptor, and retinoid X receptor. Smoking can also affect the multiple pathways including inflammatory response, DNA damaging, and oxidative stress. Reactive oxygen species (ROS) work as a second messenger in miscellaneous mitochondrial and cellular procedures and signaling pathways. On the other hand, excessive ROS might react with nucleic acid, lipids, carbohydrates, and protein causing inflammation and oxidative stress that are the main causes for the development of various metabolic disorders. Hence, this chapter will include the detailed discussion about the impact of cigarette smoking on metabolic disorder along with its effects on several enzymatic and metabolic pathways.

Background Data on household transmission of carbapenemase-producing Enterobacterales (CPE) remain limited. We studied risk of CPE household co-colonization and transmission in Ontario, Canada. Methods We enrolled CPE index cases (identified via population-based surveillance from January 2015 to October 2018) and their household contacts. At months 0, 3, 6, 9, and 12, participants provided rectal and groin swabs. Swabs were cultured for CPE until September 2017, when direct polymerase chain reaction (PCR; with culture of specimens if a carbapenemase gene was detected) replaced culture. CPE risk factor data were collected by interview and combined with isolate whole-genome sequencing to determine likelihood of household transmission. Risk factors for household contact colonization were explored using a multivariable logistic regression model with generalized estimating equations. Results Ninety-five households with 177 household contacts participated. Sixteen (9%) household contacts in 16 (17%) households were CPE-colonized. Household transmission was confirmed in 3/177 (2%) cases, probable in 2/177 (1%), possible in 9/177 (5%), and unlikely in 2/177 (1%). Household contacts were more likely to be colonized if they were the index case’s spouse (odds ratio [OR], 6.17; 95% confidence interval [CI], 1.05–36.35), if their index case remained CPE-colonized at household enrollment (OR, 7.00; 95% CI, 1.92–25.49), or if they had at least 1 set of specimens processed after direct PCR was introduced (OR, 6.46; 95% CI, 1.52–27.40). Conclusions Nine percent of household contacts were CPE-colonized; 3% were a result of household transmission. Hospitals may consider admission screening for patients known to have CPE-colonized household contacts.

We enrolled 53 consecutive in-patients with COVID-19 at six hospitals in Toronto, Canada, and tested one nasopharyngeal swab/saliva sample pair from each patient for SARS-CoV-2. Overall, sensitivity was 89% for nasopharyngeal swabs and 77% for saliva (p=NS); difference in sensitivity was greatest for sample pairs collected later in illness.

Stability studies are pivotal necessities for ensuring safety, potency, and quality of drug products throughout their shelf-life. These guarantee that pharmaceutical products will remain stable and effective under the recommended storage conditions and are considered as prerequisite for approval of any pharmaceutical product. Stability tests are routine procedures that are conducted under different conditions for investigating the effect of variation in temperature, humidity, light intensity, and time on pharmaceutical products. Hence, these studies should be conducted and evaluated according to guidelines issued by ICH (International Conference on Harmonization) or WHO. In this chapter, we have discussed the types and methods of stability tests, storage conditions, as well as ICH and WHO guidelines.

We analyzed population-based surveillance data from the Toronto Invasive Bacterial Diseases Network to describe carbapenemase-producing Enterobacteriaceae (CPE) infections during 2007-2015 in south-central Ontario, Canada. We reviewed patients' medical records and travel histories, analyzed microbiologic and clinical characteristics of CPE infections, and calculated incidence. Among 291 cases identified, New Delhi metallo-β-lactamase was the predominant carbapenemase (51%). The proportion of CPE-positive patients with prior admission to a hospital in Canada who had not received healthcare abroad or traveled to high-risk areas was 13% for patients with oxacillinase-48, 24% for patients with New Delhi metallo-β-lactamase, 55% for patients with Klebsiella pneumoniae carbapenemase, and 67% for patients with Verona integron-encoded metallo-β-lactamase. Incidence of CPE infection increased, reaching 0.33 cases/100,000 population in 2015. For a substantial proportion of patients, no healthcare abroad or high-risk travel could be established, suggesting CPE acquisition in Canada. Policy and practice changes are needed to mitigate nosocomial CPE transmission in hospitals in Canada.

Background There is mounting evidence that the recent resurgence of pertussis in many countries is in part related to the acellular vaccine, which has been administered in Canada since 1997. This vaccine elicits a different cell-mediated immune response than the previously used whole-cell vaccine, and its effectiveness wanes over time. The aim of this study is to understand the immunological, demographic and clinical factors that mediate protection from pertussis on exposure. Methods This is a household case-control study protocol. Following notification of an index case in a household, a study team will conduct a home visit to collect data and biological specimens. The study team will return to the household 8 weeks from the onset of illness in the index case. The Th1, Th2 and Th17 responses, cytokine expression, IgG subclass, blood cell counts and presence of Bordetella pertussis will be determined. We will use laboratory and statistical analyses to determine immunological differences between contacts who are infected with B. pertussis and contacts who remain healthy, and to determine which clinical and demographic covariates are associated with a reduced risk of infection. Interpretation The results of this study will be essential for understanding the immune response required for protection from infection with B. pertussis and will contribute to our understanding of the shortcomings of the current vaccine.

OBJECTIVETo measure transmission frequencies and risk factors for household acquisition of community-associated and healthcare-associated (HA-) methicillin-resistant Staphylococcus aureus (MRSA).DESIGNProspective cohort study from October 4, 2008, through December 3, 2012.SETTINGSeven acute care hospitals in or near Toronto, Canada.PARTICIPANTSTotal of 99 MRSA-colonized or MRSA-infected case patients and 183 household contacts.METHODSBaseline interviews were conducted, and surveillance cultures were collected monthly for 3 months from household members, pets, and 8 prespecified high-use environmental locations. Isolates underwent pulsed-field gel electrophoresis and staphylococcal cassette chromosome mec typing.RESULTSOverall, of 183 household contacts 89 (49%) were MRSA colonized, with 56 (31%) detected at baseline. MRSA transmission from index case to contacts negative at baseline occurred in 27 (40%) of 68 followed-up households. Strains were identical within households. The transmission risk for HA-MRSA was 39% compared with 40% (P=.95) for community-associated MRSA. HA-MRSA index cases were more likely to be older and not practice infection control measures (P=.002–.03). Household acquisition risk factors included requiring assistance and sharing bath towels (P=.001–.03). Environmental contamination was identified in 78 (79%) of 99 households and was more common in HA-MRSA households.CONCLUSIONHousehold transmission of community-associated and HA-MRSA strains was common and the difference in transmission risk was not statistically significant.Infect Control Hosp Epidemiol 2016;1–7

MicroRNAs (miRNAs) are non-coding RNAs that are associated with adipokine homeostasis and insulin resistance. Whereas, smoking can disturb metabolic homeostasis. Present study was aimed to investigate the level of miRNA-141 in experimental animal model that were exposed with graded doses of nicotine. We further aimed to investigate the possible interplay of miRNA-141 expression change with adipokine homeostasis and occurrence of insulin resistance in nicotine-exposed experimental animals. Nicotine (0.5, 1.0, 3.0 and 6.0 mg/Kg) was administered to early adolescent; postnatal days ranging from 25 to 30 Wistar rats for one month. Serum was analyzed for leptin, adipokines, IL-6, MDA, HbA1c, insulin, G6PDH, hexokinase, and lipid profile. While miRNA-141 expression level was determined in plasma. Higher doses of nicotine were associated with higher glucose, HbA1c, leptin, IL-6, MDA and lipids levels, while, insulin, adiponectin, G6PDH, hexokinase and HDL levels were lower. Higher doses of nicotine also impaired glucose tolerance and exhibited significant increase in miR-141 expression signifying that nicotine exposure may influence adipokines regulation altering glycemic profile. This is accompanied with aggravated inflammatory responses where genetic expression of miRNA-141 can be an accessible biomarker for metabolic disturbances with insulin resistance and glucose intolerance.

Background Carbapenemase-producing Enterobacteriaceae (CPE) are a global threat. Risk of transmission of CPE in households remains poorly understood Methods Population-based surveillance for CPE colonization/infection is conducted in Toronto/Peel Region, Canada. In households with ≥1 consenting household contact (HC), groin, rectal swabs and urine samples are submitted every 3 months for both IC and HC until the IC has three consecutive negative swab sets. Swabs/urines are incubated overnight in BHI, direct PCR for carbapenemase genes is performed; specimens positive for PCR are then cultured. Results Eighty-five households and 150 HC have been enrolled. Most common species/gene combinations in IC are: E. coli/NDM (33), E. coli/OXA48 (15), Klebsiella spp./NDM (11). HCs have a median of eight swabs (range 2–14). 12 (8%) HCs were colonized with CPE (median 1.5 pos samples, range 1–8). IC and HC had same gene in 11(92%) cases, and same species/gene in seven (58%) cases. NDM+OXA48 ICs were more likely to have CPE colonized HC, see table. CPE colonized HC were older, more likely to be the IC’s spouse (OR 32, 95% CI 4–260), and more likely to have travelled outside Canada (OR 9.7, 95% CI 1.2–78). Conclusion HC colonization with CPE is uncommon, but not rare, and may be associated with either household transmission, or co-exposure of HC and IC via travel. Spouses are most often colonized. Characteristic CPE Positive N = 12 CPE Negative N = 138 P-Value Gender (n, % male) 3 (25%) 53 (38%) 0.27 Median age (range) 70y(24-89) 42y (4-98) 0.005 Chronic illness 6 (50%) 35 (25%) 0.08 Relationship to IC Spouse 11(92%) 35 (25%) 3 months 7/11(64%) 32/61(52%) 0.72 >6 months 6/10(60%) 23/53(43%) 0.49 Disclosures S. Poutanen, MERCK: Scientific Advisor, Speaker honorarium; COPAN: Speaker(but not part of a bureau), Travel reimbursement; Accelerate Diagnostics: Investigator, Research support; Bio-Rad: Investigator, Research support; bioMérieux: Investigator, Research support.

Background We aimed to assess the risk of subsequent infection among patients colonized by CPE. Methods The Toronto Invasive Bacterial Diseases Network (TIBDN) has conducted population-based surveillance for CPE colonization/infection in Toronto and Peel region, Ontario, Canada, since CPE were first identified (2007). All laboratories report all CPE isolates to TIBDN. Clinical data are collected via patient interview and hospital chart review. Initially colonized patients are followed for 5y; subsequent CPE infection is defined as an episode with onset >3 days after initial detection of CPE colonization that meets National Healthcare Safety Network criteria for infection with a clinical isolate of CPE. Results From 2007 to 2018, 790 persons with CPE colonization/infection were identified. Among 364 cases colonized at identification, 42 (12%) subsequently had at least one clinical isolate, and 23 (6%) had an infection: 8 with bacteremia (primary or secondary), 7 UTI, 5 pneumonia, and 3 other. The median time from identification of colonization to infection was 21 days (IQR 7–38), with a probability of developing an infection of 7% at 3 months, and 18% by 3 years (figure). In 305 cases with data available to date, older persons, those admitted to the ICU, and those with current/recent invasive medical devices were more likely to develop infection (table). Gender, underlying conditions and other procedures were not associated with risk of infection. There was a trend to infections being more likely in patients colonized with K. pneumoniae (52% vs. 35%, P = 0.13). Conclusion The risk of subsequent infection in our cohort was 18%, with highest risk in the first 3 months; most infections occurred in patients requiring intensive care unit admission and invasive medical devices. Disclosures All authors: No reported disclosures.

Background The spread of CPE is an increasing global threat to patient safety. We describe the introduction and evolution of CPE in south-central Ontario, Canada. Methods The Toronto Invasive Bacterial Diseases Network has performed population based surveillance for CPE in metropolitan Toronto and Peel region from first identified isolates in 2007. All laboratories test/refer all carbapenem non-susceptible Enterobacterial isolates for PCR testing for carbapenemases. Demographic and medical data and travel history are collected from chart review and patient/physician interview. Results Since 2007, 659 patients have been identified as colonized/infected with CPE; 362, 57%) have at least one clinical isolate. Annual incidence has increased from 0 in 2006 to 1.3 per 100,000 in 2016/17 (Figure 1). First bacteremia occurred in 2010, the incidence in 2017 was 0.14 per 100,000 population. 388 (59%) patients were male, median age was 70 years (range 3 months–100 years). Most common genes among first isolates were NDM (306, 46%), OX48 (149, 23%), KPC (122, 19%). Most common species were K. pneumoniae (268, 41%) and E. coli (259, 39%). Over time, second species/same gene were identified in 113 (16%) patients. In addition, 34/xxx patients with isolates with NDM and/or OXA-48 subsequently had a second isolate with a different gene/gene combination. Of 518 patients whose travel and hospitalization history are available, patients with VIM were less likely than other patients to have a foreign hospitalization or travel history (9/28 vs. 341/490, P < 0.0001). Patients with KPC were more likely to have a hospitalization history outside Canada and the Indian subcontinent (25/70, 36%), in Canada (47/164,29%) than to have no hospitalization in the last year (13/93, 14%), or a history of hospitalization in the Indian subcontinent (2/191, 1%) (P < 0.001). The number of incident patients with different hospitalization and travel history over time is shown in Figure 2. Conclusion CPE is increasingly recognized in southern Ontario, both in patients with a history of exposure in healthcare in other countries, and to healthcare in Canada. Intensification of control programs is urgently needed. Disclosures S. Poutanen, MERCK: Scientific Advisor, Speaker honorarium. COPAN: Speaker(but not part of a bureau), Travel reimbursement. Accelerate Diagnostics: Investigator, Research support. Bio-Rad: Investigator, Research support. bioMérieux: Investigator, Research support.