Phylogeographical pattern of Francisella tularensis in a nationwide outbreak of tularaemia in Norway, 2011.

In 2011, a nationwide outbreak of tularaemia occurred in Norway with 180 recorded cases. It was associated with the largest peak in lemming density seen in 40 years. Francisella tularensis was isolated from 18 patients. To study the geographical distribution of F.tularensis genotypes in Norway and correlate genotype with epidemiology and clinical presentation,we performed whole genome sequencing of patient isolates. All 18 genomes from the outbreak carried genetic signatures of F. tularensis subsp. holarctica and were assigned to genetic clades using canonical single nucleotide polymorphisms. Ten isolates were assigned to major genetic clade B.6 (subclade B.7),seven to clade B.12, and one to clade B.4. The B.6 subclade B.7 was most common in southern and central Norway, while clade B.12 was evenly distributed between the southern, central and northern parts of the country. There was no association between genotype and clinical presentation of tularaemia, time of year or specimen type. We found extensive sequence similarity with F. tularensis subsp. holarctica genomes from high-endemic tularaemia areas in Sweden.Finding nearly identical genomes across large geographical distances in Norway and Sweden imply a life cycle of the bacterium without replication between the outbreaks and raise new questions about long-range migration mechanisms.

PCR-based detection and molecular characterization of shiga toxin-producing Escherichia coli strains in a routine microbiology laboratory over 16 years.

Shiga toxin-producing Escherichia coli (STEC) is a heterogeneous group of bacteria causing disease ranging from asymptomatic carriage and mild infection to hemolytic uremic syndrome (HUS). Here, we describe patients with STEC infection and characterize the STEC strains detected in our laboratory by use of PCR for stx1, stx2, and eae from 1996 through 2011. Patient information was collected from referral forms and from the Norwegian Surveillance System for Communicable Diseases. STEC isolates were characterized with respect to serogroup or serotype, selected potential virulence genes, and multilocus variable-number tandem-repeat analysis (MLVA) genotype. STEC strains were isolated from 138 (1.09%) of 12,651 patients tested. STEC strains of serogroups O26, O103, O121, O145, and O157 were the most frequent. These serogroups, except non-sorbitol-fermenting O157, were also the most frequent among the 11 patients (all ≤5 years old) who developed HUS. Twenty-four STEC strains were classified as being HUS associated based on an epidemiological link to a HUS case, including an MLVA genotype identical to that of the STEC strain. The age of the patient (≤5 years) and the genes eae and stx2a were significantly associated with HUS-associated STEC (P < 0.05 for each parameter), while stx1 was associated with non-HUS-associated STEC (P < 0.05). All of the potential virulence genes analyzed, except ehxA, were significantly more frequent among HUS-associated than non-HUS-associated strains (P < 0.05 for each gene). However, these genes were also present in some non-HUS-associated STEC strains and could therefore not reliably differentiate between HUS-associated and non-HUS-associated STEC strains.

All-time high tularaemia incidence in Norway in 2011: report from the national surveillance.

Tularaemia has mainly been a sporadic disease in Norway. In 2011, 180 persons (3.7 per 100,000 population) were diagnosed with tularaemia. This article describes the epidemiological and clinical features of tularaemia cases during a year with exceptionally high tularaemia incidence. Data from the national reference laboratory for tularaemia combined with epidemiological data from the Norwegian Surveillance System for Communicable Diseases (MSIS) were used. The incidence of tularaemia varied greatly between counties, but almost every county was involved. The majority (77.8 %) of the cases were diagnosed during the autumn and winter months. The geographic distribution also showed seasonal patterns. Overall, oropharyngeal tularaemia (41.1 %) was the most common clinical presentation, followed by glandular (14.4 %), typhoidal (14.4 %), respiratory (13.3 %) and ulceroglandular (12.8 %) tularaemia. From January to April, oropharyngeal tularaemia dominated, from May to September, ulceroglandular tularaemia was most common, whereas from October to December, there was an almost even distribution between several clinical forms of tularaemia. Eighty-five (47.2 %) of all tularaemia cases were admitted to, or seen as outpatients in, hospitals. An unexpectedly high number (3.9 %) of the patients had positive blood culture with Francisella tularensis. The clinical manifestations of tularaemia in Norway in 2011 were diverse, and changing throughout the year. Classification was sometimes difficult due to uncharacteristic symptoms and unknown mode of transmission. In rodent years, tularaemia is an important differential diagnosis to keep in mind at all times of the year for a variety of clinical symptoms.

Imported toxigenic cutaneous diphtheria in a young male returning from Mozambique to Norway, March 2014.

Six outbreaks of infectious syphilis in the United Kingdom, ongoing since 2012, have been investigated among men who have sex with men (MSM) and heterosexual men and women aged under 25 years. Interventions included case finding and raising awareness among healthcare professionals and the public. Targeting at-risk populations was complicated as many sexual encounters involved anonymous partners. Outbreaks among MSM were influenced by the use of geospatial real-time networking applications that allow users to locate other MSM within close proximity.

No increase in headache after previous intracranial infections: a historical cohort study (HUNT).

BACKGROUND AND PURPOSE: Despite the absence of robust scientific evidence, it is today generally accepted that the acute headache typical for intracranial infections can develop into permanent headache complaints. This widespread concept was explored in the first, large, longitudinal, population-based study. METHODS: Data on confirmed exposure to intracranial infections amongst all adult inhabitants in a geographical area during a 20-year period were assembled from hospital records. Surviving individuals were later invited to the third Nord-Trøndelag Health Survey (HUNT 3), where 39,690 (42%) of 94,194 invited inhabitants aged ≥20 years responded to a validated headache questionnaire. Using logistic regression, the 1-year prevalence of headache and its subtypes according to the diagnostic criteria of the International Headache Society was assessed and compared between those with and without previous confirmed intracranial infection. Age and sex were used as covariates. RESULTS: Overall, 43 participants were identified with earlier intracranial infection, whereof three had more than one infection: bacterial meningitis (n=19), lymphocytic meningitis (n=18), encephalitis (n=9), and brain abscess (n=1).The mean interval from infection to participation in HUNT 3 was 11.2 (range 1.5-19.7) years. There was no significant increase in the prevalence of headache (OR 1.10, 95% CI 0.58-2.07), its subtypes (migraine, or tension-type headache), or chronic daily headache (OR 1.85, 95% CI 0.45-7.68) amongst participants with previous intracranial infection compared with the surrounding population. CONCLUSIONS: This study challenges the existence of chronic post-bacterial meningitis headache and does not indicate the presence of other long-term headaches induced by intracranial infection.

The impact of a lung cancer computed tomography screening result on smoking abstinence.

Receiving a lung cancer computed tomography screening result might be a teachable moment for smoking cessation, but it might also unintentionally reassure smokers to continue smoking. The objective of the present study was to investigate whether test results were associated with smoking abstinence in the Dutch-Belgian Randomised Controlled Lung Cancer Screening Trial (NELSON trial). Two random samples of male smokers who had received either only negative test results (n=550) or one or more indeterminate test result (n=440) were sent a questionnaire 2 yrs after randomisation. Smokers with an indeterminate result reported more quit attempts (p=0.02), but the prolonged abstinence rate in smokers receiving a negative test (46 (8.9%) out of 519 subjects) was comparable with the abstinence rate in smokers with one or more indeterminate results (48 (11.5%) out of 419 subjects) (p=0.19). A statistically insignificant increase was found after one or more indeterminate test result (10.9 and 15.0%, respectively) compared with receiving only negative test results (8.9%) (p=0.26). In conclusion, the outcome of the screening test had no impact on future smoking abstinence in male smokers, although all results suggest more favourable implications after one or more follow-up recommendations. Screening test outcomes could be used as a teachable moment for smoking cessation.

Long-term effects of lung cancer computed tomography screening on health-related quality of life: the NELSON trial.

The long-term effects of lung cancer computed tomography (CT) screening on health-related quality of life (HRQoL) have not yet been investigated. In the Dutch-Belgian Randomised Lung Cancer Screening Trial (NELSON trial), 1,466 participants received questionnaires before randomisation (T0), 2 months after baseline screening (screen group only; T1) and at 2-yr follow-up (T2). HRQoL was measured as generic HRQoL (12-item short-form questionnaire and EuroQoL questionnaire), anxiety (Spielberger State-Trait Anxiety Inventory) and lung cancer-specific distress (impact of event scale (IES)). Repeated measures of ANOVA were used to analyse differences between the screen and control groups, and between indeterminate (requiring a follow-up CT) and negative screening result groups. At T0 and T2 there were no significant differences in HRQoL scores over time between the screen and control groups, or between the indeterminate or negative second-round screening result group. There was a temporary increase in IES scores after an indeterminate baseline result (T0: mean 4.0 (95% CI 2.8-5.3); T1: mean 7.8 (95% CI 6.5-9.0); T2: mean 4.5 (95% CI 3.3-5.8)). At 2-yr follow-up, the HRQoL of screened subjects was similar to that of control subjects, the unfavourable short-term effects of an indeterminate baseline screening result had resolved and an indeterminate result at the second screening round had no impact on HRQoL.

Outbreak of tularaemia in central Norway, January to March 2011.

From January to March 2011, 39 cases of tularaemia were diagnosed in three counties in central Norway: 21 cases of oropharyngeal type, 10 cases of glandular/ulceroglandular type, two of respiratory and two of typhoid type. Three cases were asymptomatic and clinical information was unavailable for one case. The mean age was 40.3 years (range 2-89 years). Thirty-four reported use of drinking water from private wells. An increased rodent (lemming) population and snow melting may have led to contamination of the wells with infected rodents or rodent excreta.

Short-term health-related quality of life consequences in a lung cancer CT screening trial (NELSON).

BACKGROUND: In lung cancer CT screening, participants often have an indeterminate screening result at baseline requiring a follow-up CT. In subjects with either an indeterminate or a negative result after screening, we investigated whether health-related quality of life (HRQoL) changed over time and differed between groups in the short term. METHODS: A total of 733 participants in the NELSON trial received four questionnaires: T0, before randomisation; T1, 1 week before the baseline screening; T2, 1 day after the screening; and T3, 2 months after the screening results but before the 3-month follow-up CT. HRQoL was measured as generic HRQoL (the 12-item Short Form, SF-12; the EuroQol questionnaire, EQ-5D), anxiety (the Spielberger State-Trait Anxiety Inventory, STAI-6), and lung-cancer-specific distress (the Impact of Event Scale, IES). For analyses, repeated-measures analysis of variance was used, adjusted for covariates. RESULTS: Response to each questionnaire was 88% or higher. Scores on SF-12, EQ-5D, and STAI-6 showed no clinically relevant changes over time. At T3, IES scores that were clinically relevant increased after an indeterminate result, whereas these scores showed a significant decrease after a negative result. At T3, differences in IES scores between the two baseline result groups were both significant and clinically relevant (P<0.01). CONCLUSION: This longitudinal study among participants of a lung cancer screening programme showed that in the short term recipients of an indeterminate result experienced increased lung-cancer-specific distress, whereas the HRQoL changes after a negative baseline screening result may be interpreted as a relief.

Arsenic in drinking water wells on the Bolivian high plain: Field monitoring and effect of salinity on removal efficiency of iron-oxides-containing filters.

In the rural areas around Oruro (Bolivia), untreated groundwater is used directly as drinking water. This research aimed to evaluate the general drinking water quality, with focus on arsenic (As) concentrations, based on analysis of 67 samples from about 16 communities of the Oruro district. Subsequently a filter using Iron Oxide Coated Sand (IOCS) and a filter using a Composite Iron Matrix (CIM) were tested for their arsenic removal capacity using synthetic water mimicking real groundwater. Heavy metal concentrations in the sampled drinking water barely exceeded WHO guidelines. Arsenic concentrations reached values up to 964 µ g L⁻¹ and exceeded the current WHO provisional guideline value of 10 µ g L⁻¹ in more than 50% of the sampled wells. The WHO guideline of 250 mg L⁻¹ for chloride and sulphate was also exceeded in more than a third of the samples, indicating high salinity in the drinking waters. Synthetic drinking water could be treated effectively by the IOCS- and CIM-based filters reducing As to concentrations lower than 10 µ g L⁻¹. High levels of chloride and sulphate did not influence As removal efficiency. However, phosphate concentrations in the range from 4 to 24 mg L⁻¹ drastically decreased removal efficiency of the IOCS-based filter but had no effects on removal efficiency of the CIM-based filter. Results of this study can be used as a base for further testing and practical implementation of drinking water purification in the Oruro region.

Informed participation in a randomised controlled trial of computed tomography screening for lung cancer.

The actual lung cancer (screening) knowledge, attitudes, risk perceptions, reasons to participate in or decline participation, and informed decisions of subjects who decided to or decided not to participate in the Dutch-Belgian randomised controlled trial for lung cancer screening in high-risk subjects (the NELSON trial) were evaluated. A total of 2,500 high-risk subjects were asked to complete a questionnaire 3 weeks after they had received a brochure with information about the trial. Differences in knowledge, attitude and risk perception between participants and nonparticipants were analysed with logistic regression analyses adjusted for sex and smoking status. The questionnaire response of trial participants was 80% (n = 889) whereas the response of nonparticipants was low (7%, n = 97) and selective. Participants' responses to knowledge items on lung cancer as a disease were on average more often correct (mean+/-sd 68+/-17%) than items on lung cancer screening (49+/-29%). Participants had adequate knowledge on lung cancer screening (51%) more often than the nonparticipants (38%; p = 0.009). Of the decisions regarding participation, 49% were uninformed, mainly due to insufficient knowledge. Most of the participants (99%) and 64% of the nonparticipants had a positive attitude towards lung cancer screening. Additional efforts are required to improve the knowledge and understanding of subjects who are in the process of decision-making regarding participation in a lung cancer screening trial.

Characterisation of invasive group B streptococci based on investigation of surface proteins and genes encoding surface proteins.

The joint distributions of the six genes bca, bac, epsilon/alp1, alp2, alp3 and rib (encoding alpha-C-protein, beta-C-protein, epsilon/Alp1, Alp2, Alp3, and Rib, respectively) and the proteins alpha-C-protein, beta-C-protein and Rib were investigated in invasive isolates of group B streptococcus (GBS). In total, 297 invasive isolates (123 from neonates, 174 from adults) from south-west Sweden were collected during a 13-year period. Genes were detected using multiplex and specific PCRs, and expression of the surface proteins was demonstrated using monoclonal antibodies. The genes studied were found alone or in combinations in 294 (99%) of the invasive isolates. The most common genes were rib (n = 127 isolates, 43%), alp3 (n = 78, 26%) and epsilon/alp1 (n = 42, 14%). The bac gene was never found alone, but was found in combination with one other gene in 36 isolates. The surface proteins studied were detected alone or in combinations in 152 (51%) isolates, with the most common being Rib (n = 80, 27%), alpha-C-protein (n = 68, 23%) and beta-C-protein (n = 24, 8%). Several genes were associated significantly with particular serotypes (e.g., epsilon/alp1 with serotype Ia; bca and bac with serotypes Ib and II; rib with serotype III; alp3 with serotype V). Overall, it was concluded that demonstration of different genes and surface proteins of GBS strains can be useful in epidemiological studies and in formulation of vaccines, but disappointingly, no single gene or surface protein included in the study was sufficiently common for it to be considered as the basis for a successful GBS vaccine.

Invasive group B streptococcus (GBS) disease in Norway 1996-2006.

The aim of this study was to survey the occurrence of invasive group B streptococcus (GBS) disease in Norway and detect possible trends in characteristics of invasive GBS strains from 1996 to 2006. Data from national monitoring systems for infectious diseases in Norway were analysed. Of 638,452 live births in the period, 434 cases of invasive GBS disease in infants were reported. In adults and children older than 1 year of age, 969 cases were reported. The incidence of invasive GBS disease increased significantly in the elderly, while the incidence of neonatal early-onset disease was stable with 0.46 cases per 1,000 live births. The incidence of late-onset disease increased in 2005 and 2006. The lethality of GBS in infants increased from an average of 6.5% in 1996-2005 to 20% in 2006. Serotypes III and V were predominant in 839 invasive GBS strains characterized-type III in infants and type V in the elderly. The distribution of serotypes did not change throughout the period. The distribution of detected surface proteins was stable from 1996 to 2005, but the detection rates in types III and V were low. Molecular methods for GBS typing introduced in 2006 made characterization of nearly all strains possible and appear more applicable to epidemiological studies of GBS than conventional methods. Resistance to erythromycin and clindamycin increased significantly in 2006. The increased incidence in the elderly, the increased lethality in infants in 2006, and the increased resistance to erythromycin and clindamycin the same year might indicate changing characteristics of invasive GBS strains.

Photoreactivity of biologically active compounds. XVIII. Photostability of ofloxacin in the solid state and in a tablet formulation.

The photostability of ofloxacin in the solid state has been investigated. The change in colour of uncoated and film coated ofloxacin tablets and compressed ofloxacin was studied as a function of irradiance level and total exposure energy. The degradation of ofloxacin in the various preparations was quantified by HPLC and the antimicrobial activity was determined for selected tablets. The structure of two main degradation products from ofloxacin in the solid state has been postulated from LC-MS analysis. Both products have an absorption cut-off below 400 nm and cannot explain the observed change in tablet colour. There was no apparent relationship between the change in colour and the loss of active substance or antibacterial activity for the preparations investigated. The change in colour was easily detectable at rather low exposure levels. Apparently, there was a difference in light sensitivity between the two film-coated tablet batches investigated. The results obtained were partly dependent on the conditions within the radiation chamber (e.g., exposure time and irradiance level), which emphasizes the importance of testing the samples under various conditions unless the results are unequivocal. The tablets were sensitive to visible light although ofloxacin only has a neglectible absorption above 400 nm. The film coated ofloxacin tablets did, however, absorb above 400 nm with a cut-off at approximately 520 nm. A change in tablet coating to include a component that filters visible light in addition to UV radiation might provide a solution to the discolouration problem and prevent batch to batch variations with respect to light sensitivity.

Characterization of the virulence potential of Staphylococcus condimenti isolated from a patient with severe soft tissue infection.

The coagulase-negative bacterium Staphylococcus condimenti and closely related species are commonly isolated from or found in starter cultures of fermented sausage as well as fish and soy sauces, and have traditionally been considered nonpathogenic. Recently, however, a case of catheter-related bacteraemia caused by S. condimenti was reported. In the present study we identified and characterized a strain of S. condimenti isolated from a patient with a severe soft tissue infection, comparing it to S. condimenti and S. carnosus type strains in order to elucidate the virulence potential of the clinical strain. Genome comparison showed high degree of conservation between the clinical strain and the type strain used in food industry, as well as with S. carnosus. The genome of the clinical S. condimenti strain contains few horizontally transferred regions and 37 putative virulence genes, including genes with similarity to leucocidin and genes involved in immune evasion, proinflammatory and cytolytic activity. However, it remains to be tested whether these putative virulence genes are expressed and functional. Although uncommon, S. condimenti may cause severe infection in previously healthy persons.

Streptococcus agalactiae (group B streptococcus (GBS)) colonisation and persistence, in pregnancy; a comparison of two diverse communities (rural and urban).

OBJECTIVE: To establish the extent of GBS colonisation, persistence of colonisation in pregnancy and influence of obstetric history in two diverse communities (rural and urban) in Zimbabwe. DESIGN: Cross sectional survey. SETTING: Rutsanana Clinic in Highfield, Harare (representing the urban area) and Chitsungo Mission Hospital in Lower Guruve, (representing the rural area). SUBJECTS: 300 and 100 pregnant women from the urban and rural areas respectively. MAIN OUTCOME MEASURES: GBS colonisation and persistence rates for both urban and rural areas were established, together with pregnancy outcome. RESULTS: Mother colonisation rate was significantly higher in the rural areas (60%) as compared to the urban areas (46%). GBS colonisation persistence was evidently more in rural (48%) that in urban women (12%). Baby colonisation was also more in the rural (23%) that in urban area (5%). In both the rural and urban areas, flu-like illness was a common feature and was equally reported by the subjects. Vaginal discharge requiring treatment, previous stillbirths and previous miscarriages were equally reported in both communities.

Outbreak of methicillin-resistant Staphylococcus aureus in two nursing homes in Central Norway.

Until recently, infections with methicillin-resistant Staphylococcus aureus (MRSA) have mainly been associated with hospital outbreaks in Norway. However, increasingly cases are contracted outside hospitals. This paper reports the first two outbreaks of MRSA in two nursing homes in central Norway, affecting 23 residents and five staff members. Pulsed-field gel electrophoresis analysis showed that all strains from nursing home A were identical and that the strains from nursing home B were genotypically similar with one or two band differences. Multi-locus sequence typing (MLST) showed that the strains from the two nursing homes belong to clonal complex 45, with each strain being a single-locus variant of sequence type 45 (ST 45), a well-known European epidemic strain. No evident source of the two outbreaks was found, and there was no obvious connection between the two outbreaks. The latter is also supported by the minor differences observed by MLST, suggesting a connection at some time in the past. The outbreaks led to a heavier workload and economic strain on both nursing homes. The outbreak in nursing home A was brought to an end, whereas two residents remained colonized in nursing home B despite several eradication attempts. These outbreaks show the potential for MRSA spread in a nursing home. If the prevalence of MRSA in Norway continues to increase, nursing home staff and residents may have to be included in the groups to be screened for MRSA upon hospital admission.

Ruptured abdominal aortic aneurysm secondary to tuberculous spondylitis.

A 69-year-old man was admitted with low back pain and signs of nerve root compression. A computed tomography (CT) scan showed abscess formation in the left psoas region, spondylodiscitis L3-L4 and a ruptured abdominal aortic aneurysm. The aortic aneurysm was replaced with a bifurcated vascular graft. One week later, laminectomy at the L4-level was done. In a small abscess, Mycobacterium bovis was found. The condition was considered to be a mycobacterial spondylitis secondary to BCG instillations of the urinary bladder for carcinoma. The patient received antituberculous medication for 9 months. Subsequently bone transplantation and internal fixation of the spine became necessary. Three years after surgery he is in good condition and there are no signs of graft infection on CT. Spondylitis and mycotic aortic aneurysm should be kept in mind in patients who have been treated for carcinoma of the bladder with BCG instillations.

Production of monoclonal antibodies against the human anaphylatoxin C5a des Arg and their application in the neoepitope-specific sandwich-ELISA for the quantification of C5a des Arg in plasma.

A panel of ten murine monoclonal antibodies (MAbs) was raised against the human anaphylatoxin C5a des Arg. The MAbs were shown to abrogate or significantly inhibit the chemotactic activity in zymosan activated serum. MAb 4A2E10E2 and MAb 3G3C4 were used as capture and detecting antibody, respectively, in a sandwich enzyme-linked immunosorbent assay (ELISA) for the quantification of C5a des Arg. This ELISA was shown to be very sensitive (detection limit 20 pg/ml) and could be applied directly to plasma/serum samples. The lack of interference by plasma components, in particular C5, suggested specificity for an epitope on C5a (des Arg) which is concealed in native C5 and exposed on the activation fragment only, i.e., a 'neoepitope'. The mean C5a des Arg level in EDTA-plasma from 25 healthy individuals assessed at a 1/20 dilution was 11.2 ng/ml (SD 3.4; range 6.4-16.8 ng/ml). The applicability of the assay was investigated in patients treated with haemodialysis using different membranes. Markedly elevated plasma levels of C5a des Arg were found in blood returning from the dialyzer following contact with cuprophane membranes.

Monoclonal antibodies against the major zymosan-induced chemotactic factor in rabbit serum.

The major zymosan-induced chemotactic factor in rabbit serum was purified by a two-step ion exchange chromatography procedure. The purified chemoattractant was used as antigen for production of murine monoclonal antibodies against the major chemotactic factor. The primary screening of the hybridoma cultures was an indirect ELISA using purified chemotactic factor as antigen. The final selection among ELISA-positive clones was performed according to the results obtained in a chemotactic inhibition assay. Six monoclonal antibodies were raised. These antibodies completely abrogated or substantially reduced the chemotactic activity in crude zymosan-activated serum. The chemotactic factor(s) could be absorbed onto an immunosorbent column containing monoclonal antibody and subsequently be specifically eluted with acid. By Western blot analysis the molecular weight of the major chemotactic factor was estimated to be approximately 15,000, and isoelectric focusing indicated a pI of about 9.4.