Tracing local and regional clusters of carbapenemase-producing Klebsiella pneumoniae ST512 with whole genome sequencing, Finland, 2013 to 2018.

BackgroundTwo epidemiologically-unrelated clusters of Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae were detected among several healthcare facilities (HCF) in Finland by routine surveillance using whole genome sequencing (WGS).AimThe objective was to investigate transmission chains to stop further spread of the responsible strain.MethodsIn this observational retrospective study, cases were defined as patients with K. pneumoniae KPC-3 sequence type (ST)512 strain detected in Finland from August 2013 to May 2018. Environmental specimens were obtained from surfaces, sinks and toilets in affected wards. WGS was performed on K. pneumoniae cultures using Illumina MiSeq platform and data were analysed using Ridom SeqShere software K. pneumoniae core genome multilocus sequence typing (cgMLST) scheme. Epidemiological information of the cases was provided by HCFs.ResultsWe identified 20 cases in six HCFs: cluster 1 included 18 cases in five HCFs and cluster 2 two cases in one HCF. In cluster 1, a link with a foreign country was unclear, 6/18 cases without overlapping stay had occupied the same room in one of the five HCFs within > 3 years. In cluster 2, the index case was transferred from abroad, both cases occupied the same room 8 months apart. A strain identical to that of the two cases in cgMLST was isolated from the toilet of the room, suggesting a clonal origin.ConclusionsThe clusters were mostly related to case transfer between facilities and likely involved environmental transmission. We show that CPE surveillance using WGS and collaboration between hospitals are crucial to identify clusters and trace transmission chains.

A controlled survey of less typical long-term consequences after an extensive waterborne epidemic.

BACKGROUND: Extensive backflow of treated wastewater caused household water contamination in a Finnish town in 2007. The drinking water of 9 500 residents became heavily polluted with faecal microbes, resulting in a large gastroenteritis epidemic. Cases of reactive arthritis, milder joint symptoms and prolonged gastrointestinal symptoms were observed after the outbreak. A follow-up survey was performed to study less familiar long-term health consequences within a year from the outbreak. METHODS: The contaminated group comprised a sample of residents of the area with polluted water supply (N = 323) and the control group a sample of residents in a nearby municipality (N = 186). The presence of 20 general symptoms or complaints was inquired by a mail survey. Quarterly prevalence of each symptom or complaint was measured. Twelve of these proceeded to further analysis. RESULTS: The response rate was 53% (323/615) in the contaminated group and 54% (186/343) in the control group. Rash, eye irritation, heartburn and weight loss were more prevalent in the contaminated group during the first year quarter. In the last year quarter, only eye irritation was significantly more common in the contaminated group. CONCLUSION: The excess prevalence of four complaints at the first year quarter can be explained by acute gastroenteritis or intensive water chlorination. The excess prevalence of eye irritation at the fourth year quarter cannot be explained by chlorination anymore but might be a sign of co-existing reactive joint disease. In general, long-term consequences of the outbreak can be considered minor in terms of the surveyed symptoms or complaints.

Degradation and Crystallization of Cellulose in Hydrogen Chloride Vapor for High-Yield Isolation of Cellulose Nanocrystals.

Despite the structural, load-bearing role of cellulose in the plant kingdom, countless efforts have been devoted to degrading this recalcitrant polysaccharide, particularly in the context of biofuels and renewable nanomaterials. Herein, we show how the exposure of plant-based fibers to HCl vapor results in rapid degradation with simultaneous crystallization. Because of the unchanged sample texture and the lack of mass transfer out of the substrate in the gas/solid system, the changes in the crystallinity could be reliably monitored. Furthermore, we describe the preparation of cellulose nanocrystals in high yields and with minimal water consumption. The study serves as a starting point for the solid-state tuning of the supramolecular properties of morphologically heterogeneous biological materials.

Hybrid supramolecular and colloidal hydrogels that bridge multiple length scales.

Hybrid nanocomposites were constructed based on colloidal nanofibrillar hydrogels with interpenetrating supramolecular hydrogels, displaying enhanced rheological yield strain and a synergistic improvement in storage modulus. The supramolecular hydrogel consists of naphthyl-functionalized hydroxyethyl cellulose and a cationic polystyrene derivative decorated with methylviologen moieties, physically cross-linked with cucurbit[8]uril macrocyclic hosts. Fast exchange kinetics within the supramolecular system are enabled by reversible cross-linking through the binding of the naphthyl and viologen guests. The colloidal hydrogel consists of nanofibrillated cellulose that combines a mechanically strong nanofiber skeleton with a lateral fibrillar diameter of a few nanometers. The two networks interact through hydroxyethyl cellulose adsorption to the nanofibrillated cellulose surfaces. This work shows methods to bridge the length scales of molecular and colloidal hybrid hydrogels, resulting in synergy between reinforcement and dynamics.

Modification of cellulose nanofibrils with luminescent carbon dots.

Films and hydrogels consisting of cellulose nanofibrils (CNF) were modified by covalent EDC/NHS coupling of luminescent, water-dispersible carbon dots (CDs). Quartz crystal microgravimetry with dissipation monitoring (QCM-D) and surface plasmon resonance (SPR) were used to investigate the attachment of CDs on carboxymethylated CNF (CM-CNF). As the first reported use of CD in nanocellulose products, we provide proof-of-concept for the synthesis of transparent and fluorescent nanopaper and for its tunable luminescence as confirmed by confocal microscopy imaging.

Cellulose nanocrystal-mediated synthesis of silver nanoparticles: role of sulfate groups in nucleation phenomena.

Developing sustainable chemical methods to synthesize silver nanoparticles has drawn significant research interest. Due to their unique and well-defined physical-chemical properties, cellulose nanocrystals (CNCs) have become one of the most promising renewable nanomaterials. Here we use CNC to mediate silver nanoparticle synthesis and elucidate the effect of CNC surface chemistry (as defined by sulfate groups) in nanoparticle formation and nucleation in the presence of borohydride reduction. Pristine CNCs produced by sulfuric acid hydrolysis and partially desulfated CNCs mediated the formation of silver nanoparticles of different sizes (and size distribution) following different rates of formation, as determined by transmission electron microscopy (TEM) and UV-vis spectroscopy. The results shed light on methods to stabilize silver nanoparticles, control their nucleation, and highlight the potential of CNCs in metal nanoparticle synthesis.

Adsorption of hydrophobically end-capped poly(ethylene glycol) on cellulose.

Adsorption of poly(ethylene glycol), hydrophobically end-capped with octadecenylsuccinic anhydride (OSA-PEG-OSA), on an ultrathin film of cellulose has been studied by quartz crystal microbalance with dissipation monitoring (QCM-D) and atomic force microscopy (AFM). Normally, PEG does not adsorb on cellulosic surfaces, but the use of the telechelic hydrophobic modification was found to promote adsorption. The influence of the conformation of the polymer in solution prior to adsorption and the subsequent properties of the adsorbed layer were investigated. The adsorption experiments were done at concentrations below and above the critical association concentration. The adsorption of OSA-PEG-OSA on cellulose was observed to occur in four distinct stages. Because of the amphiphilic nature of cellulose, further adsorption experiments were performed on hydrophobic (polystyrene) and hydrophilic (silica) model substrates to illuminate the contribution of hydrophobic and hydrophilic factors in the adsorption phenomenon. As expected, the kinetics and the mechanism of adsorption were strongly dependent on the chemical composition of the substrate.

Cilia-mimetic hairy surfaces based on end-immobilized nanocellulose colloidal rods.

We show a simple method toward nanoscale cilia-like structures, i.e., functional hairy surfaces, upon topochemically functionalizing nanorods of cellulose nanocrystals (CNCs) with thiol end groups (CNC-SHs), which leads to their immobilization onto a gold surface from one end, still allowing their orientational mobility. CNCs having a lateral dimension of 3-5 nm and length of 50-500 nm incorporate the native crystalline structure with hydrogen-bonded cellulose chains in the parallel configuration. This facilitates asymmetric, selective chemical modification of the reducing ends through reductive amination. Successful thiol functionalization is demonstrated using cryo transmission electron microscopy based on selective attachment of silver nanoparticles to the CNC-SH ends to form Janus-like colloidal rod-sphere adducts. The extent of thiol modification of CNC-SHs is quantified using X-ray photoelectron spectroscopy. The promoted binding of CNC-SHs on gold surfaces is shown by atomic force microscopy and quartz crystal microbalance, where the high dissipation suggests pronounced orientational mobility due to flexible joints at one rod end onto the surfaces. That the joints are flexible is also shown by the bending and realignment of the CNC-SH rods using a receding triple-phase evaporation front of a drying drop of water. The ability of the hairy surface to size-selectively resist particle binding was also investigated. As the CNCs are piezoelectric and allow magnetic functionalization by nanoparticles, we foresee a general platform for nanosized artificial cilia for fluid manipulation and controlled adsorption/desorption.

Preferential adsorption and activity of monocomponent cellulases on lignocellulose thin films with varying lignin content.

Understanding the enzymatic hydrolysis of cellulose and the influence of lignin in the process are critical for viable production of fuels and chemicals from lignocellulosic biomass. The interactions of monocomponent cellulases with cellulose and lignin substrates were investigated by using thin films supported on quartz crystal microgravimetry (QCM) resonators. Trichoderma reesei exoglucanase (CBH-I) and endoglucanase (EG-I) bound strongly to both cellulose and lignin but EG-I exhibited a distinctive higher affinity with lignin, causing a more extensive inhibition of the cellulolytic reactions. CBH-I was found to penetrate into the bulk of the cellulose substrate increasing the extent of hydrolysis and film deconstruction. In the absence of a cellulose binding domain (CBD) and a linker, the CBH-I core adsorbed slowly and was not able to penetrate into the film. Conversely to CBH-I, EG-I exhibited activity only on the surface of the lignocellulose substrate even when containing a CBD and a linker. Interestingly, EG-I displayed a clearly different interaction profile as a function of contact time registered by QCM.

Methicillin-resistant Staphylococcus aureus epidemic restricted to one health district in Finland: a population-based descriptive study in Pirkanmaa, Finland, years 2001-2011.

BACKGROUND: The incidence of methicillin-resistant Staphylococcus aureus (MRSA) has been low in the Scandinavian countries, including Finland. METHODS: We report a population-based, prospective survey of an exceptionally large ongoing MRSA epidemic in a Finnish health district (HD), Pirkanmaa HD, during 2001-2011 caused by 1 strain, spa t067 (FIN-16). RESULTS: The first FIN-16 case in the HD was identified in 2000. Ten years later, 2447 carriers had been found in 95 different institutions. MRSA in carriers was mostly health care-associated (98%). The epidemic emerged in long-term care facilities and gradually spread to acute hospitals. The majority of carriers were elderly people (median age 78 y). Fifty-two percent of new carriers had an infection with MRSA at the time of detection. CONCLUSIONS: The incidence of MRSA in Pirkanmaa HD is substantially higher than the incidence for all of Finland. Temporary projects to control MRSA have not proved sufficient to contain this large epidemic.

Clostridium difficile contamination of public tap water distribution system during a waterborne outbreak in Finland.

AIMS: In November through December 2007, the drinking water distribution system in the town of Nokia, Finland, was contaminated with treated sewage effluent that resulted in a large gastroenteritis outbreak in the community. The aim of the present study was to investigate if the contaminated water in this outbreak was also a potential source of Clostridium difficile infections. METHODS: Samples from the contaminated tap water and treated sewage effluent were collected. Stool samples from a portion of patients that fell ill during the outbreak were examined for C. difficile. PCR ribotyping was performed on toxin positive C. difficile isolates and the genetic profiles of the water and patient isolates were compared. RESULTS: Twelve toxin-positive C. difficile isolates were found in water samples: five from contaminated tap water and seven from treated sewage effluent. Among these, four and five distinct PCR ribotype profiles were identified, respectively. Four PCR ribotype profiles were found among nine human faecal C. difficile isolates. Two isolates, one from tap water and one from a patient, had an indistinguishable PCR ribotype profile. CONCLUSIONS: Our findings demonstrate for the first time C. difficile contamination of a tap water distribution system and waterborne transmission of toxigenic C. difficile seems possible.

Excess healthcare costs of a large waterborne outbreak in Finland.

BACKGROUND: The economic effects of waterborne outbreaks have rarely been reported. A large waterborne outbreak occurred in the town of Nokia in Finland in 2007 with half of the population in the contaminated area suffering from gastroenteritis. We studied the healthcare costs of this outbreak. METHODS: Healthcare costs were studied using register data from the Nokia Health Care Centre, data collected in the regional university hospital, and data from laboratory register on stool samples. RESULTS: Total excess healthcare costs were EUR 354,496, which is approximately EUR 10 per resident of Nokia. There were 2052 excess visits because of gastroenteritis in Nokia Health Care Centre, 403 excess episodes in the university hospital, and altogether over 2000 excess stool samples due to the outbreak. Care in the Nokia Health Care Centre accounted for 44% and care in the university hospital for 42% of the excess healthcare costs while stool samples accounted for only 10%. CONCLUSIONS: Despite the high morbidity, the total cost was low because most patients had a relatively mild illness. The situation would have been worse if the microbes involved had been more hazardous or if the financial situation of the community had been worse. Prevention of waterborne outbreaks is important, as there is a risk of severe short- and long-term health effects and substantial health-economic costs.

Joint symptoms after a large waterborne gastroenteritis outbreak--a controlled, population-based questionnaire study.

OBJECTIVES: Waterborne outbreaks offer an opportunity to study joint symptoms after a simultaneous exposure. In November 2007, a gastroenteritis outbreak due to faecal contamination of tap water took place in a Finnish town. The purpose of this study was to evaluate the occurrence of joint symptoms after the outbreak. METHODS: The authors conducted a controlled, population-based questionnaire survey to study the occurrence of joint symptoms within 8 weeks after the exposure. The survey covered three areas: contaminated and uncontaminated parts of the town and a control town. A total of 1000 residents were randomly selected from each area, and the joint symptoms were first analysed separately and thereafter categorized as arthritis-like if joint swelling, redness, warmth or pain in movement was reported. RESULTS: A total of 2123 responses could be evaluated. The overall prevalence of joint symptoms was 13.9% in the contaminated group, 4.3% in the uncontaminated group and 1.5% among the control group, and the frequency of arthritis-like symptoms in the groups was 6.7, 2.1 and 0.5%, respectively. Gastrointestinal symptoms predicted joint complaints, diarrhoea and blood in faeces being the most significant. Residing in the contaminated area was associated with any joint symptom [odds ratio (OR) = 4.0, 95% CI 1.8, 9.0] and joint pain (OR = 7.3, 95% CI 2.1, 24.8) without preceding gastroenteritis. CONCLUSION: The frequency of joint symptoms was high in the contaminated group and also increased in the uncontaminated group. Furthermore, the risk of joint symptoms was increased in the contaminated group even without gastroenteritis.

CMC-modified cellulose biointerface for antibody conjugation.

In this Article, we present a new strategy for preparing an antihemoglobin biointerface on cellulose. The preparation method is based on functionalization of the cellulose surface by the irreversible adsorption of CMC, followed by covalent linking of antibodies to CMC. This would provide the means for affordable and stable cellulose-based biointerfaces for immunoassays. The preparation and characterization of the biointerface were studied on Langmuir-Schaefer cellulose model surfaces in real time using the quartz crystal microbalance with dissipation and surface plasmon resonance techniques. The stable attachment of antihemoglobin to adsorbed CMC was achieved, and a linear calibration of hemoglobin was obtained. CMC modification was also observed to prevent nonspecific protein adsorption. The antihemoglobin-CMC surface regenerated well, enabling repeated immunodetection cycles of hemoglobin on the same surface.

Generic method for modular surface modification of cellulosic materials in aqueous medium by sequential "click" reaction and adsorption.

A generic approach for heterogeneous surface modification of cellulosic materials in aqueous medium, applicable for a wide range of functionalizations, is presented. In the first step, carboxymethyl cellulose (CMC) modified with azide or alkyne functionality, was adsorbed on a cellulosic substrate, thus, providing reactive sites for azide-alkyne cycloaddition click reactions. In the second step, functional units with complementary click units were reacted on the cellulose surface, coated by the click-modified CMC. Selected model functionalizations on diverse cellulosic substrates are shown to demonstrate the generality of the approach. The concept by sequentially combining the robust physical adsorption ("physical click") and robust chemical reaction ("chemical click") allows versatile, simple, and environmentally friendly modification of a cellulosic substrate with virtually any azide- or alkyne-modified molecule and even functionalization with several types of units.

Generic method for attaching biomolecules via avidin-biotin complexes immobilized on films of regenerated and nanofibrillar cellulose.

We investigated the adsorption and chemical conjugation of avidin and its deglycosylated form, neutravidin, on films of regenerated and nanofibrillar cellulose. The dynamics and extent of biomolecular attachment were monitored in situ by quartz crystal microbalance microgravimetry and ex situ via surface analyses with atomic force microscopy and X-ray photoelectron spectroscopy. The installation of carboxyl groups on cellulose after modification with carboxymethylated cellulose (CMC) or TEMPO-oxidation significantly increases physisorption of avidins, which can be then covalently conjugated by using 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride/N-hydroxysuccinimide (EDS/NHS) coupling chemistries. The developed cellulose-avidin biointerfaces are able to scavenge biotinylated molecules from solution as demonstrated by successful surface complexation of biotinylated bovine serum albumin (Biotin-BSA) and antihuman immunoglobulin G (Biotin-anti-hIgG). Finally, we show that cellulose substrates carrying immobilized anti-hIgG are effective in detecting human immunoglobulin G (hIgG) from fluid matrices.

Bicomponent lignocellulose thin films to study the role of surface lignin in cellulolytic reactions.

Ultrathin bicomponent films of cellulose and lignin derivatives were deposited on silica supports by spin coating, and after conversion into the respective polymer precursor, they were used as a model system to investigate interfacial phenomena relevant to lignocellulose biocatalysis. Film morphology, surface chemical composition, and wettability were determined by atomic force microscopy, X-ray photoelectron spectroscopy, and water contact angle, respectively. Phase separation of cellulose and lignin produced structures that resembled the cell wall of fibers and were used to monitor enzyme binding and cellulolytic reactions via quartz crystal microgravimetry. The rate and extent of hydrolysis was quantified by using kinetic models that indicated the role of the surface lignin domains in enzyme inhibition. Hydrophobic interactions between cellulases and the substrates and their critical role on irreversible adsorption were elucidated by using acetylated lignin films with different degrees of substitution. Overall, it is concluded that sensors based on the proposed ultrathin films of lignocellulose can facilitate a better understanding of the complex events that occur during bioconversion of cellulosic biomass.

Pseudoxanthomonas bacteria that drive deposit formation of wood extractives can be flocculated by cationic polyelectrolytes.

Runnability problems caused by suspended bacteria in water using industries, have, in contrast to biofilms, received little attention. We describe here that Pseudoxanthomonas taiwanensis, a wide-spread and abundant bacterium in paper machine water circuits, aggregated dispersions of wood extractives ("pitch") and resin acid, under conditions prevailing in machine water circuits (10(9) cfu ml(-1), pH 8, 45°C). The aggregates were large enough (up to 50 µm) so that they could be expected to clog wires and felts and to reduce dewatering of the fiber web. The Pseudoxanthomonas bacteria were negatively charged over a pH range of 3.2-10. Cationic polyelectrolytes of the types used as retention aids or fixatives to flocculate "anionic trash" in paper machines were effective in flocculating the Pseudoxanthomonas bacteria. The polyelectrolyte most effective for this purpose was of high molecular weight (7-8 × 10(6) g mol(-1)) and low charge density (1 meq g(-1)), whereas polyelectrolytes that effectively zeroed the electrophoretic mobility (i.e., neutralized the negative charge) of the bacterium were less effective in flocculating the bacteria. Based on the results, we concluded that the polyelectrolytes functioning by bridging mechanism, rather than by neutralization of the negative charge, may be useful as tools for reducing harmful deposits resulting from interaction of bacteria with wood extractives in warm water industry.

Sensory Appeal and Routines Beat Health Messages and Visibility Enhancements: Mixed-Methods Analysis of a Choice-Architecture Intervention in a Workplace Cafeteria.

Easier recognition and enhanced visibility of healthy options supposedly increase healthy choices, but real-world evidence remains scarce. Addressing this knowledge gap, we promoted nutritionally favourable foods in a workplace cafeteria with three choice-architectural strategies-priming posters, point-of-choice nutrition labels, and improved product placement-and assessed their effects on visual attention, food choices, and food consumption. Additionally, we developed a method for analysing real-world eye-tracking data. The study followed a pretest-posttest design whereby control and intervention condition lasted five days each. We monitored visual attention (i.e., total number and duration of fixations) and food choices with eye tracking, interviewed customers about perceived influences on food choices, and measured cafeteria-level food consumption (g). Individual-level data represents 22 control and 19 intervention participants recruited at the cafeteria entrance. Cafeteria-level data represents food consumption during the trial (556/589 meals sold). Results indicated that the posters and labels captured participants' visual attention (~13% of fixations on defined areas of interest before food choices), but the intervention had insignificant effects on visual attention to foods, on food choices, and on food consumption. Interviews revealed 17 perceived influences on food choices, the most common being sensory appeal, healthiness, and familiarity. To conclude, the intervention appeared capable of attracting visual attention, yet ineffective in increasing healthier eating. The developed method enabled a rigorous analysis of visual attention and food choices in a natural choice setting. We discuss ways to boost the impact of the intervention on behaviour, considering target groups' motives. The work contributes with a unique, mixed-methods approach and a real-world setting that enabled a multi-dimensional effects evaluation with high external validity.

Population-Based Assessment of Contact Tracing Operations for Coronavirus Disease 2019 in Pirkanmaa Hospital District, Finland.

Background: The coronavirus disease 2019 (COVID-19) epidemic overwhelmed local contact tracing (CT) efforts in many countries. In Finland, severe acute respiratory syndrome coronavirus 2 incidence and mortality were among the lowest in Europe during 2020-2021. We evaluated CT efficiency, effectiveness, and transmission settings. Methods: Polymerase chain reaction (PCR) test-positive COVID-19 cases and high-risk contacts in the population-based CT database of Pirkanmaa Hospital District (population 540 000) during June 2020-May 2021 were interviewed. Results: Altogether 353 926 PCR tests yielded 4739 (1.3%) confirmed cases (average 14-day case notification rate, 34 per 100 000 population); about 99% of confirmed cases and high-risk contacts were reached by a CT team. Of 26 881 high-risk contacts who were placed in quarantine, 2275 subsequently tested positive (48% of new cases), 825 (17%) had been in quarantine ≥48 hours before symptoms, and 3469 (77%) of locally acquired cases were part of transmission chains with an identified setting. The highest secondary attack rates were seen in households (31%), healthcare patients (18%), and private functions (10%). Among the 311 hospitalized patients, COVID-19 diagnosis or exposure was known in 273 (88%) before emergency room admission (identified patients). Healthcare workers had the highest proportion of work-related infections (159 cases [35%]). The source of infection was classifiable in 65% and was most commonly a coworker (64 cases [62%]). Conclusions: Our data demonstrate the role of effective testing and CT implementation during the cluster phase of COVID-19 spread. Although half of newly diagnosed cases were already in quarantine, targeted public health measures were needed to control transmission. CT effectiveness during widespread community transmission should be assessed.