ESBL plasmids in Klebsiella pneumoniae: diversity, transmission, and contribution to infection burden in the hospital setting

Background Resistance to third-generation cephalosporins, often mediated by extended–spectrum beta–lactamases (ESBLs), is a considerable issue in hospital-associated infections as few drugs remain for treatment. ESBL genes are often located on large plasmids that transfer horizontally between strains and species of Enterobacteriaceae and frequently confer resistance to additional drug classes. While plasmid transmission is recognised to occur in the hospital setting, the frequency and impact of plasmid transmission on infection burden, compared to ESBL+ strain transmission, is not well understood. Methods We sequenced the genomes of clinical and carriage isolates of Klebsiella pneumoniae species complex from a year long hospital surveillance study to investigate ESBL burden and plasmid transmission in an Australian hospital. Long term persistence of a key transmitted ESBL+ plasmid was investigated via sequencing of ceftriaxone resistant isolates during four years of follow–up, beginning three years after the initial study. Results We found 25 distinct ESBL plasmids. One (Plasmid A, carrying blaCTX–M–15 in an IncF backbone similar to pKPN–307) was transmitted at least four times into different Klebsiella species/lineages and was responsible for half of all ESBL episodes during the initial one-year study period. Three of the Plasmid A–positive strains persisted locally 3–6 years later, and Plasmid A was detected in two additional strain backgrounds. Overall Plasmid A accounted for 21% of ESBL+ infections in the follow–up period. Conclusions Whilst ESBL plasmid transmission events were rare in this setting, they had a significant and sustained impact on the burden of ceftriaxone resistant and multidrug resistant infections.

The use of the strain Enterococcus faecium in the technology of yogurt “Carpathian” as an ancillary culture with probiotic properties

The results of research on the technological features of the production of yogurt "Carpathian" are covered in the article. The bacterial preparation of Chr. Hansen series YoFlex Premium 1.0 (L. bulgaricus, S. thermophilus) and Creamy 1.0 (L. bulgaricus, S. thermophilus, L. rhamnosus) and strain E. faecium SB 18, which is isolated from traditional Carpathian fermented products were used to produce research yogurt samples. It was found that when the strains were used together, the microorganisms were compatible, did not show interspecific antagonism and did not inhibit the enzymatic process. Based on yogurt microorganisms and E. faecium SB 18 strain, seven prototypes of yogurt were created: № 1 (100 %) – control, Premium + Creamy; № 2 (100 %) – control, pure culture of E. faecium SB 18; №3 (100:100 %) – control, (Premium + Creamy) + E. faecium SB 18; №4 (50:50 %) – (Premium + Creamy) + Ent. faecium SB 18; № 5 (70:30 %) – (Premium + Creamy) + Ent. faecium SB 18; № 6 (80:20 %) – (Premium + Creamy) + Ent. faecium SB 18; № 7 (90:10 %) – (Premium + Creamy) + Ent. faecium SB 18. The fastest fermentation took place in sample № 1 (pH 4.78 units – 4 h), the slowest in sample №2 (pH 4.81 units – 6 h), where only pure culture of E. faecium SB 18 was used. The fermentation time in sample №3 was initially slower and then more active (pH 4.77 units – 4 h). The acidity increased more moderately in samples № 4, 5, 6, 7 for 4 h, and at the end of fermentation was 4.84 units, 4.76 units, 4.81 units. and 4.75 units. in accordance. According to organoleptic evaluation, the experimental samples were characterized by slight differences. In general, it is noted that the addition of microbial culture of E. faecium SB 18 improves the taste of yogurt. Samples № 6 and № 7 with the addition of E. faecium SB 18 strain in the amount of 20 and 10 % were noted for the best organoleptic properties. According to the score scale, the above-mentioned samples received the highest number of points – 48, out of a possible 50. The dependence of the acidity of yogurt during storage was established on the dose and composition of the bacterial preparation. It was investigated that the acidity of yogurt, which contained an additional strain of E. faecium SB 18 in the ratios of 100:100 (sample 3) and 50:50 (sample 4), tends to increase rapidly in acidity, which is associated with increased lactic acid bacteria. It was found that partial replacement of the amount of traditional yogurt leaven in the ratio of 70:30 (sample 5), 80:20 (sample 6) and 90:10 (sample 7) provides the optimal course of the enzymatic process during fermentation and storage. It was found that the use of traditional yogurt leaven YoFlex Premium 1.0 and Creamy 1.0. together with strain E. faecium SB 18 in a ratio of 80:20, provides excellent consumer properties of the product.

EP.WE.213Short-term impact of COVID-19 on surgical services in a tertiary hospital

Aim Since December 2019, SARS-CoV-2 has dramatically impacted the global landscape. One of the biggest challenges has been the additional strain put on healthcare systems. Although there are numerous studies on the effects of COVID-19 on intensive care beds and ventilator availability, there has been little exploration into the wider impacts of COVID-19 on the provision of other services. This study was designed to explore how COVID-19 has impacted surgical service provision at a large NHS hospital. Methods We compared the number and types of general surgical procedures carried out in a tertiary centre in the six months prior to the UK COVID-19 outbreak (September 2019-February 2020) and the six months after (March 2020-August 2020). Results We found that since March 2020 there has been a 70% decrease in the amount of operations taking place, with numbers dropping from a pre-COVID total of 1761 to a post-COVID total of 529. This mainly affected elective procedures with emergency surgeries remaining relatively constant (48 pre-COVID vs 44 post-COVID). Conclusion COVID-19 has caused a significant decrease in the number of surgeries being undertaken. This is due to a combination of factors including staffing issues, reduced investigative capacity, and national mandates on the cessation of non-urgent procedures. Although this mainly affected elective operations, it will have wider implications on future NHS workload and training. The knock on effects will inevitably result in a rise in delayed and emergency presentations with worse patient outcomes.

Taylor, the Disability Disclosure Virtual Assistant: A Case Study of Participatory Research with Disabled Students

Administrative burden in education is a serious issue for disabled students. Form-filling and bureaucracy are ubiquitous in further and higher education, particularly for students who need to disclose a disability and arrange for accommodations and support for an equitable educational experience. Paradoxically, many of these processes are inherently inaccessible for disabled students, and yet completing them can be critical to their success. Artificial Intelligence has potential to alleviate some of the burden imposed by administration and bureaucracy; virtual assistants and chatbots can replace forms with dialogue, without placing additional strain on institutions. However, it is essential that solutions are designed in partnership with disabled students to ensure that students’ needs are met, their concerns addressed, and the final solution is equitable for them. This paper explores a case study of participatory research with disabled students in a large UK distance learning institution, in which participatory research identified an issue of administrative burden for disabled students, and a virtual assistant was designed as a solution using participatory design. It shares the methodology and design process, explores findings from different phases of the research, and shares recurrent themes arising throughout the study. In doing so, it aims to provide a foundation for future participatory research to reduce barriers for disabled students.

Escherichia coli AraJ boosts utilization of arabinose in metabolically engineered cyanobacterium Synechocystis sp. PCC 6803

Lignocellulosic biomass can serve as an inexpensive and renewable source of carbon for the biosynthesis of commercially important compounds. L-arabinose is the second most abundant pentose sugar present in the plant materials. Model cyanobacterium Synechocystis sp. PCC 6803 is incapable of catabolism of L-arabinose as a source of carbon and energy. In this study, all the heterologous genes expressed in Synechocystis were derived from Escherichia coli K-12. Initially we constructed four Synechocystis strains that expressed AraBAD enzymes involved in L-arabinose catabolism, either in combination with or without one of the three arabinose transporters, AraE, AraFGH or AraJ. Among the recombinants, the strain possessing AraJ transporter was observed to be the most efficient in terms of dry biomass production and L-arabinose consumption. Later, an additional strain was generated by the expression of AraJ in the AraE-possessing strain. The resultant strain was shown to be advantageous over its parent. This study demonstrates that AraJ, a protein with hitherto unknown function plays a role in the uptake of L-arabinose to boost its catabolism in the transgenic Synechocystis strains. The work also contributes to the current knowledge regarding metabolic engineering of cyanobacteria for the utilization of pentose sugars.

Building on the Foundation of a Sustainable Hand Hygiene Program During the COVID-19 Pandemic

Background: Hand hygiene is essential to preventing the spread of disease in hospitals. Renewed emphasis has been placed on hand hygiene during the COVID-19 pandemic. We investigated whether UNC Medical Center’s well-established Clean-In Clean-Out (CICO) program for hand hygiene observations was sustainable throughout a public health and healthcare crisis and whether the COVID-19 pandemic had an effect on hand hygiene compliance. Methods: UNC Medical Center utilizes a crowd-sourced hand-hygiene audit application, CICO, to track hand-hygiene observations, compliance, and feedback. This application encourages participation from all staff and promotes providing real-time feedback in the form of a compliment or reminder when performing hand hygiene observations. During this evaluation, hand hygiene data were queried from the CICO application on the number of observations performed, hand hygiene compliance percentage, and feedback compliance percentage from July 2019 to December 2020. Hand hygiene data were compared to patient volumes in different care settings and the number of hospitalized patients being treated for COVID-19. Results: Initial increases in hand hygiene observations, compliance, and feedback were detected in the months leading up to UNC Medical Center receiving its first SARS-CoV-2–positive patient. Observations were highest when patient volumes were low due to closed clinics and restrictions on elective surgeries (Figure 1). When patient volumes returned to pre–COVID-19 levels coupled with treating more COVID-19 patients, the number of observations and compliance rate metrics declined. Feedback compliance percentage remained relatively stable through the entire period (Figure 2). Conclusions: Despite the additional strain on healthcare staff during COVID-19, the CICO model was a sustainable method to track hand hygiene observations and compliance. Notably, however, engagement was highest when patient census was lower, demonstrating that operating at a high capacity is not beneficial for patient safety. Due to the success and sustainment of the CICO program, UNC Medical Center used this model to create a Mask-On Mask-Up campaign to engage staff to submit observations, track compliance, and encourage feedback to promote the appropriate use of masks during COVID-19.Funding: NoDisclosures: None

Patterns of endogenous and exogenous ovarian hormone modulation on recovery metrics across the menstrual cycle

IntroductionAs the number of female athletes competing rises globally, training methodologies should reflect sex differences across critical metrics of adaptation to training. Surrogate markers of the autonomic nervous system (ANS) used for monitoring training load are heart rate variability (HRV) and resting heart rate (RHR). The aim was to investigate ovarian hormone effects on standard recovery metrics (HRV, RHR, respiratory rate (RR) and sleep duration) across a large population of female athletes.MethodsA retrospective study analysed 362 852 days of data representing 13 535 menstrual cycles (MC) from 4594 respondents (natural MC n=3870, BC n=455, progestin-only n=269) for relationships and/or differences between endogenous and exogenous ovarian hormones on ANS.ResultsHRV and return to baseline (recovery) decreased as resting HR and RR increased (p<0.001) from the early follicular to the late luteal phase of the MC. Patterning was paradoxical across phases for users of combined hormonal contraception (BC) as compared with the patterning of the MC. HRV and recovery start elevated and drop off quickly during the withdrawal bleed, rising through the active pill weeks (p<0.001). Progestin-only users had similar patterning as the MC. The relationship between normalised recovery and previous day strain is modulated by birth control type. BC exhibited steeper declines in recovery with additional strain-normalised recovery decreases by an additional 0.0055±0.00135 (p<0.001) per unit of strain; with no significant difference between MC and progestin-only (p=0.19).ConclusionThe patterning of ANS modulation from ovarian hormones is significantly different between naturally cycling women and those on BC, with the patterning dependent on the type of contraception used.

A mixed methods protocol to evaluate the effectiveness and acceptability of COVID-19 Community Assessment Hubs

Background: Ireland’s health system has been under significant strain due to staff shortages and inadequate capacity. Critical care bed capacity per capita in Ireland is among the lowest in Europe, thus, the coronavirus disease 2019 (COVID-19) pandemic has put additional strain on an over-stretched system. COVID-19 Community Assessment Hubs (CAHs) were established to mitigate unnecessary admission to acute hospitals, and reduce infection spread by supporting COVID-19 positive or suspected positive patients to isolate at home, or in isolation facilities. There is some evidence that similar assessment centres may be a successful triage strategy to reduce burden on hospital and acute care. Aim : The aim of this study is to evaluate the impact of COVID-19 Community Assessment Hubs on service delivery in two regions in Ireland during the pandemic. Methods: A mixed-methods approach will be used, incorporating co-design to engage stakeholders and ensure informed data capture and analysis. Online surveys will assess CAH patients’ experiences of access to and quality of care. Clinical patient data from CAHs will be collected and analysed using multinomial logistic regression to check for association with patient demographics and COVID-19 symptoms, and CAH early warning scores and outcomes (Transfer to Emergency Department, Transfer to isolation unit, Sent home with care plan). Semi-structured interviews will be conducted with: patients to elicit an in-depth understanding of experiences and acceptability of attending CAHs; and staff to understand challenges, benefits, and effectiveness of CAHs. Interview data will be analysed using thematic analysis. Discussion: This study will provide valuable insights from both patient and staff perspectives on the operation of CAHs. We will evaluate the effectiveness and acceptability of CAHs and propose areas for improvement of the service. This will contribute to international literature on the use of community assessment centres during infectious disease pandemics.

Continuity of routine immunization programs in Canada during the COVID-19 pandemic

IntroductionIn Canada, the COVID-19 pandemic has interrupted many routine health services, placed additional strain on the health care system, and resulted in many Canadians being either unable or unwilling to attend routine immunization appointments. We sought to capture and synthesize information about changes to routine immunization programs in response to the pandemic and plans to catch-up any missed immunizations.MethodsProvincial/territorial (P/T) public health leaders were interviewed via teleconference between August-October 2020 to collect information on the following topics: how routine immunization delivery was affected during and after initial lockdown periods, plans to catch-up missed doses, and major challenges and achievements in continuing routine immunization programs. Data were coded and categorized according to common responses and descriptive analysis was performed.ResultsInterviews occurred with participants from 11 of 13 P/Ts. School immunization programs were reported to be most negatively affected by the pandemic (n=9). In the early pandemic period, infant, preschool, and maternal/prenatal programs were prioritized, with most P/Ts continuing these services with adaptations for COVID-19. After the initial lockdown period, all routine programs were continuing with adaptations in most P/Ts. Infant, preschool, and school programs were most often targeted for catch-up through measures such as appointment rebooking and making additional clinics and/or providers available. Major challenges included resource limitations (e.g., staff shortages, PPE shortages, limited infrastructure) (n=11), public health restrictions (n=8), and public hesitancy to attend appointments (n=5).ConclusionsCanadian routine immunization programs faced some disruptions due to the COVID-19 pandemic, particularly the school, adult, and older adult programs. Further research is needed to determine the measurable impact of the pandemic on routine vaccine coverage levels.

Personalized and Automated Feedback in Summative Assessment Using Recommender Systems

In this study we explore the use of recommender systems as a means of providing automated and personalized feedback to students following summative assessment. The intended feedback is a personalized set of test questions (items) for each student that they could benefit from practicing with. Recommended items can be beneficial for students as they can support their learning process by targeting specific gaps in their knowledge, especially when there is little time to get feedback from instructors. The items are recommended using several commonly used recommender system algorithms, and are based on the students' scores in a summative assessment. The results show that in the context of the Dutch secondary education final examinations, item recommendations can be made to students with an acceptable level of model performance. Furthermore, it does not take a computationally complex model to do so: a simple baseline model which takes into account global, student-specific, and item-specific averages obtained similar performance to more complex models. Overall, we conclude that recommender systems are a promising tool for helping students in their learning process by combining multiple data sources and new methodologies, without putting additional strain on their instructors.