Comparative Risk Analysis of Low Back Pain among Professional Football, Ice Hockey, and Floorball Athletes.

BACKGROUND Low back pain (LBP) is a common concern among professional athletes, potentially hindering performance and career longevity. However, comparative assessments of LBP prevalence and severity across various sports remain scarce. This study aimed to evaluate the factors associated with LBP in 388 professional athletes, including football, ice hockey, and floorball players. MATERIAL AND METHODS Conducted from June 2021 to September 2022, this cross-sectional study incorporated 388 athletes from national elite clubs, including football (n=148), ice hockey (n=179), and floorball (n=61). The Oswestry Disability Index (ODI), comprising sections like pain intensity, self-care, lifting, walking, sitting, standing, sleeping, sexual life, social life, and traveling, was employed to evaluate spinal pain and disability. RESULTS The study found no significant disparities in the LBP assessment among the groups. The relative risk (OR) of LBP and disability varied among the sports: football players displayed a lower risk (OR=0.49; 95% CI 0.32-0.74, P≤0.001), while ice hockey players had a higher risk (OR=2.18; 95% CI 1.45-3.29, P≤0.001) compared to the others. In contrast, the risk for floorball players (OR=0.82; 95% CI 0.47-1.41) did not significantly deviate from that of the other two sports. CONCLUSIONS LBP prevalence stood at 42.6% for football players, 60.1% for ice hockey players, and 49.2% for floorball players. Among these, ice hockey players exhibited a 2.18-fold increased risk of developing LBP and associated disability when compared to their football and floorball counterparts.

Genomic epidemiology of Mycobacterium bovis infection in sympatric badger and cattle populations in Northern Ireland.

Bovine tuberculosis (bTB) is a costly, epidemiologically complex, multi-host, endemic disease. Lack of understanding of transmission dynamics may undermine eradication efforts. Pathogen whole-genome sequencing improves epidemiological inferences, providing a means to determine the relative importance of inter- and intra-species host transmission for disease persistence. We sequenced an exceptional data set of 619 Mycobacterium bovis isolates from badgers and cattle in a 100 km2 bTB 'hotspot' in Northern Ireland. Historical molecular subtyping data permitted the targeting of an endemic pathogen lineage, whose long-term persistence provided a unique opportunity to study disease transmission dynamics in unparalleled detail. Additionally, to assess whether badger population genetic structure was associated with the spatial distribution of pathogen genetic diversity, we microsatellite genotyped hair samples from 769 badgers trapped in this area. Birth death models and TransPhylo analyses indicated that cattle were likely driving the local epidemic, with transmission from cattle to badgers being more common than badger to cattle. Furthermore, the presence of significant badger population genetic structure in the landscape was not associated with the spatial distribution of M. bovis genetic diversity, suggesting that badger-to-badger transmission is not playing a major role in transmission dynamics. Our data were consistent with badgers playing a smaller role in transmission of M. bovis infection in this study site, compared to cattle. We hypothesize, however, that this minor role may still be important for persistence. Comparison to other areas suggests that M. bovis transmission dynamics are likely to be context dependent, with the role of wildlife being difficult to generalize.

Streptococcus canis genomic epidemiology reveals the potential for zoonotic transfer.

Streptococcus canis, a multi-host pathogen commonly isolated from dogs and cats, has been occasionally reported in severe cases of human infection. To address the gap in knowledge on its virulence and host tropism, we investigated S. canis genomic epidemiology and report the results of this analysis for the first time. We analysed 59 S. canis whole genome sequences originating from a variety of host species, comprising 39 newly sequenced isolates from UK sources, along with all (n=20) publicly available genomes. Antimicrobial resistance (AMR) phenotype was determined for all 39 available isolates. Genomes were screened for determinants of resistance and virulence. We created a core SNP phylogeny and compared strain clustering to multi-locus sequence typing (MLST) and S. canis M-like protein (SCM) typing. We investigated the dataset for signals of host adaptation using phylogenetic analysis, accessory genome clustering and pan-genome-wide association study analysis. A total of 23 % (9/39) of isolates exhibited phenotypic resistance to lincosamides, macrolides and/or tetracyclines. This was complemented by the identification of AMR-encoding genes in all genomes: tetracycline (tetO 14 %, 8/59; and tetM 7 %, 4/59) and lincosamide/macrolide (ermB, 7 %, 4/59). AMR was more common in human (36 %, 4/11) compared to companion animal (18 %, 5/28) isolates. We identified 19 virulence gene homologues, 14 of which were present in all strains analysed. In an S. canis strain isolated from a dog with otitis externa we identified a homologue of S. pyogenes superantigen SMEZ. The MLST and SCM typing schemes were found to be incapable of accurately representing core SNP-based genomic diversity of the S. canis population. No evidence of host adaptation was detected, suggesting the potential for inter-species transmission, including zoonotic transfer.

Streptococcus canis, the underdog of the genus.

Streptococcus canis is a multi-host pathogen that causes disease of varying severity in a wide range of mammals, including humans. Dogs and cats appear to be the primary hosts and may play a role in transmitting infection to humans. The broader epidemiology of S. canis, however, is still poorly understood, as are its virulence mechanisms, antimicrobial resistance (AMR) and population structure. In this review we gather existing knowledge on S. canis, describing its epidemiology in animals and humans and present information on virulence factors, classification schemes and AMR prevalence. We describe the main ecological niches of S. canis in companion animals, discuss potential risk factors for infection in humans and propose a multi-host transmission cycle. We show that current knowledge on S. canis virulence determinants is limited and sometimes contradictory. We illustrate the different typing systems proposed to classify S. canis. We also report the range of known AMR phenotypes and the emergence of new mechanisms of resistance. Finally, we discuss the zoonotic potential of S. canis, highlighting the need for further evidence in this area. Streptococcus canis may be regarded as a neglected pathogen of one health concern. Further research is needed for its better understanding and effective control.

Target-enrichment sequencing yields valuable genomic data for challenging-to-culture bacteria of public health importance.

Genomic data contribute invaluable information to the epidemiological investigation of pathogens of public health importance. However, whole-genome sequencing (WGS) of bacteria typically relies on culture, which represents a major hurdle for generating such data for a wide range of species for which culture is challenging. In this study, we assessed the use of culture-free target-enrichment sequencing as a method for generating genomic data for two bacterial species: (1) Bacillus anthracis, which causes anthrax in both people and animals and whose culture requires high-level containment facilities; and (2) Mycoplasma amphoriforme, a fastidious emerging human respiratory pathogen. We obtained high-quality genomic data for both species directly from clinical samples, with sufficient coverage (>15×) for confident variant calling over at least 80% of the baited genomes for over two thirds of the samples tested. Higher qPCR cycle threshold (Ct) values (indicative of lower pathogen concentrations in the samples), pooling libraries prior to capture, and lower captured library concentration were all statistically associated with lower capture efficiency. The Ct value had the highest predictive value, explaining 52 % of the variation in capture efficiency. Samples with Ct values ≤30 were over six times more likely to achieve the threshold coverage than those with a Ct > 30. We conclude that target-enrichment sequencing provides a valuable alternative to standard WGS following bacterial culture and creates opportunities for an improved understanding of the epidemiology and evolution of many clinically important pathogens for which culture is challenging.

The hospital environment versus carriage: transmission pathways for third-generation cephalosporin-resistant bacteria in blood in neonates in a low-resource country healthcare setting.

Neonatal bloodstream infections (BSI) can lead to sepsis, with high morbidity and mortality, particularly in low-income settings. The high prevalence of third-generation cephalosporin-resistant organisms (3GC-RO) complicates the management of BSI. Whether BSI is linked to carriage of 3GC-RO, or to acquisition from the hospital environment is important for infection prevention and control, but the relationship remains unclear, especially in low-income settings. At a tertiary hospital in Mwanza, Tanzania, we screened neonatal blood and rectal samples from 200 neonates, and 400 (hospital) environmental samples. We used logistic regression to identify risk factors, and Kolmogorov-Smirnov tests and randomisation analyses to compare distributions of species and resistance patterns to assess potential routes of transmission. We found that BSIs caused by 3GC-RO were frequent (of 59 cases of BSI, 55 were caused by 3GC-RO), as was carriage of 3GC-RO, particularly Escherichia coli, Klebsiella pneumoniae, and Acinetobacter species. In the 28 infants with both a carriage and blood isolate, there were more (4 of 28) isolate pairs of the same species and susceptibility profile than expected by chance (p < 0.05), but most pairs were discordant (24 of 28). Logistic regression models found no association between BSI and carriage with either 3GC-RO or only 3GC-R K. pneumoniae. These analyses suggest that carriage of 3GC-RO is not a major driver of BSI caused by 3GC-RO in this setting. Comparison with environmental isolates showed very similar distributions of species and resistance patterns in the carriage, BSI, and the environment. These similar distributions, a high frequency of Acinetobacter spp. isolations, the lack of strong association between carriage and BSI, together with the high proportion of 3GC-RO in BSI all suggest that these neonates acquire multidrug-resistant carriage and blood isolates directly from the hospital environment.

Pelvic floor muscle training, the risk of falls and urgency urinary incontinence in older women.

PURPOSE: The primary aim of the work was to evaluate the effect of physical training and pelvic floor muscle training combined with dual tasks (PFMT-DT) in older women on urgency urinary incontinence (UUI). The secondary aim of the work was to evaluate the impact of interventions on static and dynamic balance, risk of falls and fear of falls. SAMPLE: The research sample consisted of 88 older women with UUI, with a mean age of 75 ± 4.3 years. They were randomly divided into two groups: the experimental (n = 40) and control (n = 40) groups. METHODS: The duration of the intervention was 12 weeks. Both groups underwent physical training three times a week for 30 minutes. In addition, the experimental group received pelvic floor muscle training in different positions and with dual cognitive tasks two times a week for 30 minutes. We used The International Consultation on Incontinence Questionnaire (ICIQ-UI SF), the modified Voiding Diary, and the Overactive Bladder Questionnaire(OAB-q). The risk of falls were assessed according to Tinetti's Performance Oriented Mobility Assessment. Fear of falls was assessed by the Falls Efficacy Scale. RESULTS: After the treatment, significant differences between groups were recorded in favor of the experimental group in the daytime frequency of voiding (7.6 to 5.3), in nycturia (2.1 to 0.7), in UUI (1.8 to 1.0), OAB-q SS (40.8 to 17.6) and OAB-q HR (61.2 to 83.8) (p ≤ 0.001), with large effect size (ES), as well as in the Tinetti balance and gait and in the fall risk assessment (19.2 to 23.2) (p ≤ 0.001), also with a large ES. For fear of falls, significant differences were noted (80.0 to 71.5) (p ≤ 0.05), with a small ES in favor of the experimental group. CONCLUSION: The PFMT-DT proved to be an effective intervention in improving the symptoms of OAB and UUI. We were able to significantly reduce the risk of falls according to POMA by about 21% (19.2 ± 2.7 to 23.20 ± 3.25%) in older women with UUI.

Long-term maintenance of multidrug-resistant Escherichia coli carried by vampire bats and shared with livestock in Peru.

Extended-spectrum beta-lactamase-producing Escherichia coli (ESBL-E. coli) have been reported in wildlife worldwide. Whether wildlife is a transient host of ESBL-E. coli or comprises an independently maintained reservoir is unknown. We investigated this question by longitudinally monitoring ESBL-E. coli in common vampire bats and nearby livestock in Peru. Among 388 bats from five vampire bat colonies collected over three years, ESBL-E. coli were detected at a low prevalence (10% in 2015, 4% in 2017 and 2018) compared to a high prevalence (48%) from 134 livestock sampled in 2017. All ESBL-E. coli were multidrug-resistant, and whole genome sequencing of 33 randomly selected ESBL-E. coli isolates (18 recovered from bats) detected 46 genes conferring resistance to antibiotics including third-generation cephalosporins (e.g., blaCTX-M-55, blaCTX-M-15, blaCTX-M-65, blaCTX-M-3, blaCTX-M-14), aminoglycosides, fluoroquinolones, and colistin (mcr-1). The mcr-1 gene is reported for the first time on a wild bat in Latin America. ESBL-E. coli also carried 31 plasmid replicon types and 16 virulence genes. Twenty-three E. coli sequence types (STs) were detected, including STs involved in clinical infections worldwide (e.g., ST 167, ST 117, ST 10, ST 156 and ST 648). ESBL-E. coli with identical cgMLST (ST 167) were detected in the same bat roost in 2015 and 2017, and several ESBL-E. coli from different bat roosts clustered together in the cgMLST reconstruction, suggesting long-term maintenance of ESBL-E. coli within bats. Most antibiotic resistance and virulence genes were detected in E. coli from both host populations, while ESBL-E. coli ST 744 was found in a bat and a pig from the same locality, suggesting possible cross-species exchanges of genetic material and/or bacteria between bats and livestock. This study suggests that wild mammals can maintain multidrug-resistant bacteria and share them with livestock.

Genomic epidemiology of clinical ESBL-producing Enterobacteriaceae in a German hospital suggests infections are primarily community- and regionally-acquired.

Clinical Enterobacteriaceae isolates that produce extended-spectrum ß-lactamases (ESBLs) have been increasingly reported at a global scale. However, comprehensive data on the molecular epidemiology of ESBL-producing strains are limited and few studies have been conducted in non-outbreak situations.We used whole-genome sequencing to describe the population structure of 294 ESBL-producing Escherichia coli and Klebsiella pneumoniae isolates that were recovered from a German community hospital throughout a 1 year sampling period in a non-outbreak situation.We found a high proportion of E. coli isolates (61.5 %) belonged to the globally disseminated extraintestinal pathogenic ST131, whereas a wider diversity of STs was observed among K. pneumoniae isolates. The E. coli ST131 population in this study was shaped by multiple introductions of strains as demonstrated by contextual genomic analysis including ST131 strains from other geographical sources. While no recent common ancestor of the isolates of the current study and other international isolates was found, our clinical isolates clustered with those previously recovered in the region. Furthermore, we found that the isolation of ESBL-producing clinical strains in hospitalized patients could only rarely be associated with likely patient-to-patient transmission, indicating primarily a community and regional acquisition of strains.Further genomic analyses of clinical, carriage and environmental isolates is needed to uncover hidden transmissions and thus discover the most common sources of ESBL-producing pathogen infections in our hospitals.

Developing practical clinical tools for predicting neonatal mortality at a neonatal intensive care unit in Tanzania.

BACKGROUND: Neonatal mortality remains high in Tanzania at approximately 20 deaths per 1000 live births. Low birthweight, prematurity, and asphyxia are associated with neonatal mortality; however, no studies have assessed the value of combining underlying conditions and vital signs to provide clinicians with early warning of infants at risk of mortality. The aim of this study was to identify risk factors (including vital signs) associated with neonatal mortality in the neonatal intensive care unit (NICU) in Bugando Medical Centre (BMC), Mwanza, Tanzania; to identify the most accurate generalised linear model (GLM) or decision tree for predicting mortality; and to provide a tool that provides clinically relevant cut-offs for predicting mortality that is easily used by clinicians in a low-resource setting. METHODS: In total, 165 neonates were enrolled between November 2019 and March 2020, of whom 80 (48.5%) died. We competed the performance of GLMs and decision trees by resampling the data to create training and test datasets and comparing their accuracy at correctly predicting mortality. RESULTS: GLMs always outperformed decision trees. The best fitting GLM showed that (for standardised risk factors) temperature (OR 0.61, 95% CI 0.40-0.90), birthweight (OR 0.33, 95% CI 0.20-0.52), and oxygen saturation (OR 0.66, 95% CI 0.45-0.94) were negatively associated with mortality, while heart rate (OR 1.59, 95% CI 1.10-2.35) and asphyxia (OR 3.23, 95% 1.25-8.91) were risk factors. To identify the tool that balances accuracy and with ease of use in a low-resource clinical setting, we compared the best fitting GLM with simpler versions, and identified the three-variable GLM with temperature, heart rate, and birth weight as the best candidate. For this tool, cut-offs were identified using receiver operator characteristic (ROC) curves with the optimal cut-off for mortality prediction corresponding to 76.3% sensitivity and 68.2% specificity. The final tool is graphical, showing cut-offs that depend on birthweight, heart rate, and temperature. CONCLUSIONS: Underlying conditions and vital signs can be combined into simple graphical tools that improve upon the current guidelines and are straightforward to use by clinicians in a low-resource setting.

Complete Genome Sequences of Three Invasive Strains of Streptococcus pyogenes Subtype emm5.23 Isolated in Scotland.

Streptococcus pyogenes emm5.23 is uncommon; however, it has recently been involved in a relatively high proportion of cases of invasive disease in Scotland. Here, we report the complete genome sequences of three emm5.23 isolates, which may be used as a reference for investigating the virulence and epidemiology of this strain.

Existence of Multiple ESBL Genes among Phenotypically Confirmed ESBL Producing Klebsiella pneumoniae and Escherichia coli Concurrently Isolated from Clinical, Colonization and Contamination Samples from Neonatal Units at Bugando Medical Center, Mwanza, Tanzania.

The proportions and similarities of extended-spectrum ß-lactamase (ESBL) producing K. pneumoniae (ESBL-KP) and E. coli (ESBL-EC) carrying multiple ESBL genes is poorly known at our setting. This study investigated the existence of multiple ESBL genes (blaCTX-M, blaTEM, and blaSHV) among ESBL-KP and ESBL-EC concurrently isolated from clinical, colonization, and contamination samples from neonatology units in Mwanza-Tanzania. Twenty and 55 presumptive ESBL-EC and ESBL-KP, respectively, from a previous study archived at -80 °C were successfully recovered for this study. Isolates were screened and confirmed for production of ESBLs by phenotypic methods followed by multiplex PCR assay to determine ESBL genes. All (100%) and 97.3% of presumptive ESBL isolates were phenotypically confirmed by Clinical and Laboratory Standards Institute (CLSI) and modified double-disc synergy methods, respectively. About 93.3% (70/75) of phenotypically confirmed ESBL isolates had at least one ESBL gene, whereby for 62.9% (44/70), all ESBL genes (blaCTX-M, blaTEM, and blaSHV) were detected. Eight pairs of ESBL bacteria show similar patterns of antibiotics susceptibility and ESBL genes. ESBL-KP and ESBL-EC, concurrently isolated from clinical, colonization and contamination samples, harbored multiple ESBL genes. Further, eight pairs of ESBL isolates had similar patterns of antibiotics susceptibility and ESBL genes, suggesting transmission of and/or sharing of mobile genetic elements (MGEs) among ESBL-KP and ESBL-EC.

Bacteremia in critical care units at Bugando Medical Centre, Mwanza, Tanzania: the role of colonization and contaminated cots and mothers' hands in cross-transmission of multidrug resistant Gram-negative bacteria.

BACKGROUND: Multidrug resistance (MDR) is a major clinical problem in tertiary hospitals in Tanzania and jeopardizes the life of neonates in critical care units (CCUs). To better understand methods for prevention of MDR infections, this study aimed to determine, among other factors, the role of MDR-Gram-negative bacteria (GNB) contaminating neonatal cots and hands of mothers as possible role in transmission of bacteremia at Bugando Medical Centre (BMC), Mwanza, Tanzania. METHODS: This cross-sectional, hospital-based study was conducted among neonates and their mothers in a neonatal intensive care unit and a neonatology unit at BMC from December 2018 to April 2019. Blood specimens (n = 200) were sub-cultured on 5% sheep blood agar (SBA) and MacConkey agar (MCA) plates. Other specimens (200 neonatal rectal swabs, 200 maternal hand swabs and 200 neonatal cot swabs) were directly inoculated on MCA plates supplemented with 2 µg/ml cefotaxime (MCA-C) for screening of GNB resistant to third generation cephalosporins, r-3GCs. Conventional biochemical tests, Kirby-Bauer technique and resistance to cefoxitin 30 µg were used for identification of bacteria, antibiotic susceptibility testing and detection of MDR-GNB and screening of potential Amp-C beta lactamase producing GNB, respectively. RESULTS: The prevalence of culture confirmed bacteremia was 34.5% of which 85.5% were GNB. Fifty-five (93.2%) of GNB isolated from neonatal blood specimens were r-3GCs. On the other hand; 43% of neonates were colonized with GNB r-3GCs, 32% of cots were contaminated with GNB r-3GCs and 18.5% of hands of neonates' mothers were contaminated with GNB r-3GCs. The prevalences of MDR-GNB isolated from blood culture and GNB r-3GCs isolated from neonatal colonization, cots and mothers' hands were 96.6, 100, 100 and 94.6%, respectively. Significantly, cyanosis (OR[95%CI]: 3.13[1.51-6.51], p = 0.002), jaundice (OR[95%CI]: 2.10[1.07-4.14], p = 0.031), number of invasive devices (OR[95%CI]: 2.52[1.08-5.85], p = 0.031) and contaminated cot (OR[95%CI]: 2.39[1.26-4.55], p = 0.008) were associated with bacteremia due to GNB. Use of tap water only (OR[95%CI]: 2.12[0.88-5.09], p = 0.040) was protective for bacteremia due to GNB. CONCLUSION: High prevalence of MDR-GNB bacteremia and intestinal colonization, and MDR-GNB contaminating cots and mothers' hands was observed. Improved cots decontamination strategies is crucial to limit the spread of MDR-GNB. Further, clinical presentations and water use should be considered in administration of empirical therapy whilst awaiting culture results.

Tuberculosis bacillary load, an early marker of disease severity: the utility of tuberculosis Molecular Bacterial Load Assay.

In this comparative biomarker study, we analysed 1768 serial sputum samples from 178 patients at 4 sites in Southeast Africa. We show that tuberculosis Molecular Bacterial Load Assay (TB-MBLA) reduces time-to-TB-bacillary-load-result from days/weeks by culture to hours and detects early patient treatment response. By day 14 of treatment, 5% of patients had cleared bacillary load to zero, rising to 58% by 12th week of treatment. Fall in bacillary load correlated with mycobacterial growth indicator tube culture time-to-positivity (Spearmans r=-0.51, 95% CI (-0.56 to -0.46), p<0.0001). Patients with high pretreatment bacillary burdens (above the cohort bacillary load average of 5.5log10eCFU/ml) were less likely to convert-to-negative by 8th week of treatment than those with a low burden (below cohort bacillary load average), p=0.0005, HR 3.1, 95% CI (1.6 to 5.6) irrespective of treatment regimen. TB-MBLA distinguished the bactericidal effect of regimens revealing the moxifloxacin-20 mg rifampicin regimen produced a shorter time to bacillary clearance compared with standard-of-care regimen, p=0.008, HR 2.9, 95% CI (1.3 to 6.7). Our data show that the TB-MBLA could inform clinical decision making in real-time and expedite drug TB clinical trials.

A Tuberculosis Molecular Bacterial Load Assay (TB-MBLA).

Tuberculosis is caused by Mycobacterium tuberculosis (Mtb), a pathogen classified by the United Nations (UN) as a dangerous category B biological substance. For the sake of the workers' safety, handling of all samples presumed to carry Mtb must be conducted in a containment level (CL) 3 laboratory. The TB molecular bacterial load assay (TB-MBLA) test is a reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) test that quantifies Mtb bacillary load using primers and dual-labelled probes for 16S rRNA. We describe the use of heat inactivation to render TB samples noninfectious while preserving RNA for the TB-MBLA. A 1 mL aliquot of the sputum sample in tightly closed 15 mL centrifuge tubes is boiled for 20 min at either 80 °C, 85 °C, or 95 °C to inactivate Mtb bacilli. Cultivation of the heat inactivated and control (live) samples for 42 days confirmed the death of TB. The inactivated sample is then spiked with 100 µL of the extraction control and RNA is extracted following the standard RNA isolation procedure. No growth was observed in the cultures of heat treated samples. The isolated RNA is subjected to real-time RT-qPCR, which amplifies a specific target in the Mtb 16S rRNA gene, yielding results in the form of quantification cycles (Cq). A standard curve is used to translate Cq into bacterial load, or estimated colony forming units per mL (eCFU/mL). There is an inverse relationship between Cq and the bacterial load of a sample. The limitation is that heat inactivation lyses some cells, exposing the RNA to RNases that cause a loss of <1 log10eCFU/mL (i.e., <10 CFU/mL). Further studies will determine the proportion of very low burden patients that cause false negative results due to heat inactivation.

Biomarkers of health and welfare: A One Health perspective from the laboratory side.

A biomarker is any measurement reflecting an interaction between a biological system and a potential hazard, which may be chemical, physical, or biological. The One World, One Health concept established that human and animal health and the environmental state are highly interconnected, sharing common aspects that can be applied globally in these three components. In this paper, we review how the concept of One Health can be applied to biomarkers of health and welfare, with a special focus on five points that can be applied to any biomarker when it is expected to be used to evaluate the human, animal or environmental health. Three of these points are: (1) the different biomarkers that can be used, (2) the different sample types where the biomarkers can be analysed, and (3) the main methods that can be used for their measurement. In addition, we will evaluate two key points needed for adequate use of a biomarker in any situation: (4) a proper analytical validation in the sample that it is going to be used, and (5) a correct selection of the biomarker. It is expected that this knowledge will help to have a broader idea about the use of biomarkers of health and welfare and also will contribute to a better and more accurate use of these biomarkers having in mind their One Health perspective.

The Microevolution and Epidemiology of Staphylococcus aureus Colonization during Atopic Eczema Disease Flare.

Staphylococcus aureus is an opportunistic pathogen and variable component of the human microbiota. A characteristic of atopic eczema (AE) is colonization by S. aureus, with exacerbations associated with an increased bacterial burden of the organism. Despite this, the origins and genetic diversity of S. aureus colonizing individual patients during AE disease flares is poorly understood. To examine the microevolution of S. aureus colonization, we deep sequenced S. aureus populations from nine children with moderate to severe AE and 18 non-atopic children asymptomatically carrying S. aureus nasally. Colonization by clonal S. aureus populations was observed in both AE patients and control participants, with all but one of the individuals carrying colonies belonging to a single sequence type. Phylogenetic analysis showed that disease flares were associated with the clonal expansion of the S. aureus population, occurring over a period of weeks to months. There was a significant difference in the genetic backgrounds of S. aureus colonizing AE cases versus controls (Fisher exact test, P = 0.03). Examination of intra-host genetic heterogeneity of the colonizing S. aureus populations identified evidence of within-host selection in the AE patients, with AE variants being potentially selectively advantageous for intracellular persistence and treatment resistance.

Mycobacterial Load Assay.

Tuberculosis is a difficult disease to treat, a process made more harder as tools to monitor treatment response only provide a result long after the patient has provided a sample. The mycobacterial load assay (MBLA) provides a simple molecular test to quantify and determine the viability of M. tuberculosis in human or other samples.

Phenotypic resistance in mycobacteria: is it because I am old or fat that I resist you?

OBJECTIVES: We aimed to explore the phenomenon of phenotypic resistance to antimycobacterial antibiotics and to determine whether this was associated with cell age or the presence of lipid bodies. METHODS: The accumulation of lipid-body-positive [lipid-rich (LR)] cells was followed using cell staining and flow cytometry. LR cells of Mycobacterium smegmatis, Mycobacterium marinum, Mycobacterium fortuitum and Mycobacterium bovis (BCG) were separated from non-lipid-body-containing [lipid-poor (LP)] cells and their MBCs determined. We also compared the MBCs for LR and LP cells from 'old' and 'young' cultures. RESULTS: The LR cells of all species were more resistant to antibiotics than LP cells. For BCG, the susceptibility ratios were as follows: rifampicin, 5×; isoniazid, 16.7×; ethambutol, 5×; and ciprofloxacin, 5×. Phenotypic resistance was found in LR cells irrespective of cell age. CONCLUSIONS: We have shown that phenotypic antibiotic resistance is associated with the presence of lipid bodies irrespective of cell age. These data have important implications for our understanding of relapse in mycobacterial infections.

Genomic Investigations unmask Mycoplasma amphoriforme, a new respiratory pathogen.

BACKGROUND: Mycoplasma amphoriforme has been associated with infection in patients with primary antibody deficiency (PAD). Little is known about the natural history of infection with this organism and its ability to be transmitted in the community. METHODS: The bacterial load was estimated in sequential sputum samples from 9 patients by quantitative polymerase chain reaction. The genomes of all available isolates, originating from patients in the United Kingdom, France, and Tunisia, were sequenced along with the type strain. Genomic data were assembled and annotated, and a high-resolution phylogenetic tree was constructed. RESULTS: By using high-resolution whole-genome sequencing (WGS) data, we show that patients can be chronically infected with M. amphoriforme manifesting as a relapsing-remitting bacterial load, interspersed by periods when the organism is undetectable. Importantly, we demonstrate transmission of strains within a clinical environment. Antibiotic resistance mutations accumulate in isolates taken from patients who received multiple courses of antibiotics. CONCLUSIONS: Mycoplasma amphoriforme isolates form a closely related species responsible for a chronic relapsing and remitting infection in PAD patients in the United Kingdom and from immunocompetent patients in other countries. We provide strong evidence of transmission between patients attending the same clinic, suggesting that screening and isolation may be necessary for susceptible patients. This work demonstrates the critical role that WGS can play in rapidly unraveling the biology of a novel pathogen.