Cortinarius subgenus Leprocybe, unexpected diversity and significant differences in species compositions between western and eastern North America.

The focus of this paper is the North American species of Cortinarius in subg. Leprocybe. Eighteen species, including twelve new ones, and two tentative (aff.) species, are delimited based on morphological and molecular data (DNA ITS-LSU sequences). Existing type specimens of species in subg. Leprocybe were also studied, and neo- or epitypes designated for C. cotoneus, C. melanotus, C. phrygianus and C. venetus to stabilize the nomenclature. In addition, to improve the infrasubgeneric classification of Leprocybe three new sections are proposed: sect. Fuscotomentosi, sect. Melanoti and sect. Squamiveneti. This study adds substantial information to the knowledge of subg. Leprocybe in North America against a background of European species. To date only two species, C. phrygianus and C. squamivenetus have been reported from both continents. Citation: Ammirati J, Liimatainen K, Bojantchev D, et al. 2021. Cortinarius subgenus Leprocybe, unexpected diversity and significant differences in species compositions between western and eastern North America. Persoonia 46: 216-239. https://doi.org/10.3767/persoonia.2021.46.08.

External quality assessment for molecular typing of Salmonella 2013-2015: performance of the European national public health reference laboratories.

We report the results of three consecutive External Quality Assessments (EQAs) for molecular subtyping of Salmonella to assess the performance of the European national public health reference laboratories (NPHRLs). The EQA included the molecular typing methods used for European enhanced surveillance of human Salmonella infections: pulsed field gel electrophoresis (PFGE), including gel analysis by the use of the software BioNumerics, and 5-locus multiple locus variable number of tandem repeat analysis (MLVA) for serovar Typhimurium. The participation in the PFGE laboratory part was higher (27/35) than in the gel analysis (19/35) and MLVA (15/35), suggestive of the need for capacity building in methods requiring specialized equipment (MLVA) or software (gel analysis). The majority (25/27) of the participating NPHRLs produced inter-laboratory comparable PFGE gel(s). Two laboratories continued to produce low-quality gels and should have additional technical assistance in the future. In particular, two gel quality evaluation parameters, measuring "image acquisition and running conditions" and "bands", were identified to cause gel quality problems throughout the EQAs. Despite the high number of laboratories participating in the PFGE laboratory part, the participation in gel analysis was low, although increasing. In the MLVA part, the NPHRLs correctly assigned 96% (405/420) allelic profiles according to the nomenclature. In conclusion, the EQAs identified critical parameters for unsuccessful performance and helped to offer assistance to those laboratories that needed it most. The assessments supported the development of quality in molecular typing and promoted the harmonization of subtyping methods used for EU/EEA-wide surveillance of human Salmonella infections.

Cortinarius section Bicolores and section Saturnini (Basidiomycota, Agaricales), a morphogenetic overview of European and North American species.

Cortinarius is the largest genus of ectomycorrhizal fungi worldwide. Recent molecular studies have shown high levels of morphological homoplasy within the genus. Importantly, DNA phylogenies can reveal characteristics that have been either over- or underemphasized in taxonomic studies. Here we sequenced and phylogenetically analysed a large set of pan-European and North American collections taxonomically studied and placed in Cortinarius sect. Bicolores and sect. Saturnini, according to traditional morpho-anatomical criteria. Our goal was to circumscribe the evolutionary boundaries of the two sections, to stabilize both the limits and nomenclature of relevant species, and to identify described taxa which, according to our current understanding, belong to other lineages. Our analysis resolves two clades: /Bicolores, including 12 species, one of which is new to science, and /Saturnini, including 6 species. Fifteen binomials, traditionally treated in these two sections based on morphology, do not belong to the above two phylogenetic clades. Instead, six of these latter are clearly placed in other clades that represent sect. Bovini, sect. Sciophylli, sect. Duracini and sect. Brunneotincti. The presence or absence of blue pigments and the detection of specific odours emerge as clearly misleading taxonomic features, but more surprisingly, spore size and ecology can be misleading as well. A total of 63 type specimens were sequenced, 4 neotypes and 2 epitypes are proposed here, and 1 new combination is made.

Food-borne diseases associated with frozen berries consumption: a historical perspective, European Union, 1983 to 2013.

Epidemiological investigations of outbreaks of hepatitis A virus (HAV) and norovirus (NoV) infections in the European Union/European Economic Area (EU/EEA) in the last five years have highlighted frozen berries as a vehicle of infection. Given the increasing berry consumption in the EU over the last decades, we undertook a review of the existing evidence to assess the potential scale of threat associated with this product. We searched the literature and four restricted-access online platforms for outbreak/contamination events associated with consumption of frozen berries. We performed an evaluation of the sources to identify areas for improvement. The review revealed 32 independent events (i.e. outbreak, food contamination) in the period 1983­2013, of which 26 were reported after 2004. The identified pathogens were NoV, HAV and Shigella sonnei. NoV was the most common and implicated in 27 events with over 15,000 cases reported. A capture­recapture analysis was performed including three overlapping sources for the period 2005­2013. The study estimated that the event-ascertainment was 62%. Consumption of frozen berries is associated with increasing reports of NoV and HAV outbreaks and contamination events, particularly after 2003. A review of the risks associated with this product is required to inform future prevention strategies. Better integration of the available communication platforms and databases should be sought at EU/EEA level to improve monitoring, prevention and control of food-borne-related events.

The largest type study of Agaricales species to date: bringing identification and nomenclature of Phlegmacium (Cortinarius) into the DNA era.

Cortinarius is a species-rich and morphologically challenging genus with a cosmopolitan distribution. Many names have not been used consistently and in some instances the same species has been described two or more times under separate names. This study focuses on subg. Phlegmacium as traditionally defined and includes species from boreal and temperate areas of the northern hemisphere. Our goals for this project were to: i) study type material to determine which species already have been described; ii) stabilize the use of Friesian and other older names by choosing a neo- or epitype; iii) describe new species that were discovered during the process of studying specimens; and iv) establish an accurate ITS barcoding database for Phlegmacium species. A total of 236 types representing 154 species were studied. Of these 114 species are described only once whereas 40 species had one ore more synonyms. Of the names studied only 61 were currently represented in GenBank. Neotypes are proposed for 21 species, and epitypes are designated for three species. In addition, 20 new species are described and six new combinations made. As a consequence ITS barcodes for 175 Cortinarius species are released.

Raw grated beetroot linked to several outbreaks of sudden-onset gastrointestinal illness, Finland 2010.

In 2010, 7/44 (16%) reported foodborne outbreaks in Finland were linked with raw beetroot consumption. We reviewed data from the national outbreak registry in order to hypothesize the aetiology of illness and to prevent further outbreaks. In the seven outbreaks, 124 cases among 623 respondents were identified. Consumption of raw beetroot was strongly associated with gastrointestinal illness (relative risk 8∙99, 95% confidence interval 6∙06-13∙35). The illness was characterized by sudden onset of gastrointestinal symptoms; the median incubation time was 40 min and duration of illness 5 h. No common foodborne pathogens or toxins were found in either clinical or beetroot samples, but all tested beetroot samples were of poor quality according to total bacterial counts. Beta-haemolytic Pseudomonas fluorescens was detected in several beetroot samples but its effect on human health is unknown. No outbreaks were reported after the Finnish Food Safety Authority Evira advised against serving raw beetroot in institutional canteens.

Laboratory preparedness for detection and monitoring of Shiga toxin 2-producing Escherichia coli O104:H4 in Europe and response to the 2011 outbreak.

A hybrid strain of enteroaggregative and Shiga toxin 2-producing Escherichia coli (EAEC-STEC) serotype O104:H4 strain caused a large outbreak of haemolytic uraemic syndrome and bloody diarrhoea in 2011 in Europe. Two surveys were performed in the European Union (EU) and European Economic Area (EEA) countries to assess their laboratory capabilities to detect and characterise this previously uncommon STEC strain. Prior to the outbreak, 11 of the 32 countries in this survey had capacity at national reference laboratory (NRL) level for epidemic case confirmation according to the EU definition. During the outbreak, at primary diagnostic level, nine countries reported that clinical microbiology laboratories routinely used Shiga toxin detection assays suitable for diagnosis of infections with EAEC-STEC O104:H4, while 14 countries had NRL capacity to confirm epidemic cases. Six months after the outbreak, 22 countries reported NRL capacity to confirm such cases following initiatives taken by NRLs and the European Centre for Disease Prevention and Control (ECDC) Food- and Waterborne Disease and Zoonoses laboratory network. These data highlight the challenge of detection and confirmation of epidemic infections caused by atypical STEC strains and the benefits of coordinated EU laboratory networks to strengthen capabilities in response to a major outbreak.

A phase I, dose-escalation study of TB-403, a monoclonal antibody directed against PlGF, in patients with advanced solid tumours.

BACKGROUND: TB-403 (RO 5323441), a humanised monoclonal antibody, is a novel antiangiogenesis agent directed against placental growth factor. The safety, pharmacokinetics (PK), and antitumour activity of TB-403 were assessed in a phase I, dose-escalation study in patients with advanced solid tumours. METHODS: Patients in sequential dose groups received either weekly doses of 1.25, 5.0, or 10 mg kg(-1) or doses of 20 or 30 mg kg(-1) every third week. RESULTS: Twenty-three patients were enrolled and received TB-403. The most common adverse events (AEs) were fatigue, constipation, pyrexia, dyspnoea, and nausea. One serious AE, a lung embolus in a patient with non-small cell lung cancer treated with 10 mg kg(-1) weekly, was deemed possibly related to TB-403. No dose-limiting toxicities were observed, and a maximum-tolerated dose was not reached. The PK parameters were dose linear and the terminal half-life values ranged from 9 to 14 days. Six patients exhibited stable disease for at least 8 weeks. Two patients, (oesophageal squamous cell carcinoma and pancreatic adenocarcinoma) both treated with 5 mg kg(-1) weekly, remained stable for 12 months. CONCLUSION: TB-403 treatment in this patient population is well tolerated, with a safety profile distinct from that of vascular endothelial growth factor-axis inhibitors.

Multiple norovirus outbreaks linked to imported frozen raspberries.

In 2009, the number of foodborne norovirus outbreaks in Finland seemed markedly high, and many outbreaks seemed to be linked to imported frozen raspberries. We reviewed the data regarding all notified foodborne outbreaks in 2009 in Finland in order to assess the magnitude of the problem and to summarize the information on raspberry-linked outbreaks. Between March and August, 13 norovirus outbreaks affecting about 900 people could be linked to imported frozen raspberries. Two raspberry samples corresponding to two batches of raspberries were positive for norovirus. These two batches proved to have been the likely source in six of the 13 outbreaks. Analytical studies had not been conducted for six outbreaks, and virological test results were inconclusive in two. However, combining epidemiological and microbiological methods often enabled finding the source, as exemplified in investigation of a large school outbreak. To ensure prompt control measures in similar situations in the future, both aspects of outbreak investigations should be strengthened.

Detection of human norovirus from frozen raspberries in a cluster of gastroenteritis outbreaks.

We describe a cluster of norovirus outbreaks affecting about 200 people in Southern Finland in September and October 2009. All outbreaks occurred after consumption of imported raspberries from the same batch intended for the catering sector. Human norovirus genotype GI.4 was found in frozen raspberries. The berries were served in toppings of cakes in separate catering settings or mixed in curd cheese as a snack for children in a daycare center. The relative risk for consumption of the berry dish was 3.0 (p

Botulism and hot-smoked whitefish: a family cluster of type E botulism in France, September 2009.

A family cluster of three cases of type E botulism were identified in south-east France in September 2009. The suspected food source of infection was a vacuum packed hot-smoked whitefish of Canadian origin purchased by the family during a visit to Finland and consumed several weeks later in France on the day prior to symptom onset. No leftover fish was available to confirm this hypothesis. Vacuum packed hot-smoked whitefish has previously been associated with cases of type E botulism in multiple countries, including Finland, Germany, the United States and Israel.

Risk ranking of pathogens in ready-to-eat unprocessed foods of non-animal origin (FoNAO) in the EU: initial evaluation using outbreak data (2007-2011).

Foods of non-animal origin (FoNAO) are consumed in a variety of forms, being a major component of almost all meals. These food types have the potential to be associated with large outbreaks as seen in 2011 associated with VTEC O104. In order to identify and rank specific food/pathogen combinations most often linked to human cases originating from FoNAO in the EU, a semi-quantitative model was developed using seven criteria: strength of associations between food and pathogen based on the foodborne outbreak data from EU Zoonoses Monitoring (2007-2011), incidence of illness, burden of disease, dose-response relationship, consumption, prevalence of contamination and pathogen growth potential during shelf life. The top ranking food/pathogen combination was Salmonella spp. and leafy greens eaten raw followed by (in equal rank) Salmonella spp. and bulb and stem vegetables, Salmonella spp. and tomatoes, Salmonella spp. and melons, and pathogenic Escherichia coli and fresh pods, legumes or grains. Despite the inherent assumptions and limitations, this risk model is considered a tool for risk managers, as it allows ranking of food/pathogen combinations most often linked to foodborne human cases originating from FoNAO in the EU. Efforts to collect additional data even in the absence of reported outbreaks as well as to enhance the quality of the EU-specific data, which was used as input for all the model criteria, will allow the improvement of the model outputs. Furthermore, it is recommended that harmonised terminology be applied to the categorisation of foods collected for different reasons, e.g. monitoring, surveillance, outbreak investigation and consumption. In addition, to assist future microbiological risk assessments, consideration should be given to the collection of additional information on how food has been processed, stored and prepared as part of the above data collection exercises.

Different Yersinia enterocolitica 4:O3 genotypes found in pig tonsils in Southern Germany and Finland.

The distribution of different genotypes of Y. enterocolitica 4:O3 strains recovered from pig tonsils in Southern Germany and Finland in 1999-2000 was investigated. A total of 96 and 207 Y. enterococolitica 4:O3 isolates recovered from 47 and 66 tonsils of finishing pigs in Germany and Finland, respectively, were characterised with PFGE using NotI enzyme. In all, 39 different NotI profiles were obtained, only one of which, NB1, was found in both Germany and Finland. All strains were further characterised with ApaI and XhoI enzymes. When the 54 German and 74 Finnish strains were characterised with all three enzymes, 51 genotypes were obtained. The 23 genotypes found in German strains differed from the 28 found in Finnish strains. These results indicate that Y. enterocolitica 4:O3 genotypes have a differential geographical distribution and thus can be used in epidemiological studies.

Diffractive optical element based sensor for surface quality inspection of concave punches.

Optical surface quality of concave shaped punches was investigated with a diffractive optical element based sensor. Image information of the present sensor was studied for the purpose of surface roughness and reflectance inspection. The robust sensor is proposed for off-line optical inspection of concave punches.

Dependence between dissolution rate and porosity of compressed erythromycin acistrate tablets.

The correlation between dissolution rate and porosity of compressed erythromycin acistrate tablets was studied. The total porosity of the tablets, the pore size distribution and the specific surface area of the pores were determined using high-pressure mercury porosimetry. The particle size and specific surface area of the raw material and of the dry granulated mass of the tablets were also determined. The results show that the pore size distribution, showing the differences in pore structure, is more informative than total intruded volume of mercury. However, it is very difficult to explain the dissolution behaviour of erythromycin acistrate tablets only by porosity results of the tablets, and more work is still needed in this field.

The effect of compression force on surface structure, crushing strength, friability and disintegration time of erythromycin acistrate tablets.

The surface roughness of erythromycin acistrate tablets was studied by non-contact laser profilometry. Seven roughness parameters and 3D fractal dimension were examined. The mechanical properties (including crushing strength, friability and disintegration time) were determined, and SEM data were taken from the tablets. According to the results, the crushing strength and the disintegration time of the tablets increased with increasing compression force. At higher compression forces the crushing strength reached a constant level. The friability of the tablets behaved quite unexpectedly and minimum friability was observed at a compression force of 14 kN. Except for fractal dimension, the roughness parameters behaved very much in the same way as the friability of the tablets. The SEM data supported the friability and surface roughness data of the tablets.

Optical inspection of punches: flat surfaces.

Optical inspection techniques for estimation of surface roughness and surface quality of flat punches, which are used in the pharmaceutical industry for tablet compression, were studied. It was observed that a simple specular reflection technique is useful for surface roughness assessment of flat punches. A diffractive element based sensor was used for the surface quality detection, such as mirror property or flatness, with the aid of image information of a test pattern. It was observed that a new punch may have a poorer surface quality than a used one.

Characterisation of sucrose-negative Yersinia enterocolitica 4/O:3 isolates recovered from pig tonsils.

Sucrose-negative Yersinia enterocolitica isolates of bioserotype 4/O:3 have been recovered for the first time. They were found in 2% of the tonsils of clinically healthy fattening pigs. These sucrose-negative Y. enterocolitica isolates could not be differentiated from Y. kristensenii isolates using API 20E; thus, they were identified using PCR and sequencing. Using pulsed-field gel electrophoresis (PFGE). NotI profiles of sucrose-negative Y. enterocolitica 4/O:3 isolates showed a high similarity to sucrose-positive Y. enterocolitica 4/O:3 isolates. This study demonstrated that sucrose-negative Y. enterocolitica 4/O:3 isolates of porcine origin can harbour virulence genes; plasmid-encoded virulence markers were found in 8 out of 11 isolates and all isolates contained chromosomal-encoded virulence markers. Thus, the pathogenicity of sucrose-negative Yersinia isolates should always be assessed.

Assessing the safety environment in work organization of road maintenance jobs.

The present study examines workers' responses to the safety environment in relation to organizational, individual, and situational factors in road maintenance jobs. A self-administered questionnaire including 80 variables was used to collect data from the workers. The study population consisted of 207 workers at nine work places. The survey response rate was 93% (193 workers). The attitudes of coworkers, and judgments and attentiveness had a statistically highly significant effect on the worker's own attitude. The attitudes of supervisors, attitudes of coworkers, and the manner of instructing had a statistically highly significant effect on the performance feedback. The workers felt that the establishment of safe work habits would be most affected by the road supervisor's management methods and feedback. A considerable proportion of the workers believed that risk taking was part of their job.