Vomnedbrytningsprofil av fiber i helsäd

Syftet med detta projekt var att studera inverkan av gröda och skördetidpunkt på kemisk sammansättning och vomnedbrytning av neutral detergent fibre (NDF) och torrsubstans (TS) i helsäd samt att jämföra ANKOM DaisyII in vitro-metoden med in situ-metoden med avseende på nedbrytningsförloppet i vommen av NDF och TS i helsäd. Havre, korn, rågvete och vårvete odlades i storrutor med tre upprepningar i fält. Hela försöket upprepades under två år (2002 och 2003). Varje storruta delades upp i två smårutor, som skördades vid tidig mjölkmognad respektive tidig degmognad. In vitro-inkubering skedde med ANKOM DaisyII-inkubator av 48 prover från båda åren. Prover vägdes in i filterpåsar som förseglades och inkuberades i vomvätska och buffert i 4-liters glasflaskor, där samtliga prover för en tid fanns i samma flaska, i värmeskåp. In situ-inkubering skedde av 24 prover endast från första odlingsåret. Prov vägdes in i nylonpåsar som placerades i vommen på tre fistulerade kor. Vid både in situ- och in vitro-metoden avbröts inkuberingarna vid olika tidsintervall upp till 120 timmar genom att påsarna sköljdes i vatten. Fiberdata från in situ- och in vitro-inkuberingarna anpassades till en icke-linjär modell för skattning av potentiellt nedbrytbar NDF, nedbrytningshastigheten av NDF, potentiellt icke nedbrytbar NDF och tidsfördröjning innan nedbrytning av NDF började. Havre och vårvete innehöll mest NDF, acid detergent fibre (ADF) och lignin (P < 0,001). Korn innehöll mer NDF än rågvete. Halten NDF och ADF minskade med senare utvecklingsstadium (P < 0,001) samtidigt som stärkelsehalten ökade (P < 0,001). Ligninhalten påverkades inte av utvecklingsstadium. Korn hade den högsta stärkelsehalten följt av havre och vårvete (P < 0,001). Smältbarheten av organisk substans (OS) var högst i korn och rågvete enligt både den svenska VOS-metoden (vomvätskelöslig organisk substans) och den danska metoden EFOS (enzymfordøjeligt organisk stof; P < 0,001). Smältbarheten av OS enligt EFOS ökade med senare utvecklingsstadium (P < 0,001) medan VOS endast visade en ökning för rågvete (P < 0,05). Variationen i smältbarhet av OS enligt EFOS kunde till stor del förklaras av variationer i innehåll av stärkelse (R2=0,40), ADF (R2=0,88) och NDF (R2=0,71). Korn och rågvete gav högre TS-försvinnande (löslig + nedbrytbar TS) vid inkuberingen än havre och vårvete första året både in situ och in vitro (P < 0,001). Andra året hade korn högre TS-försvinnande in vitro än rågvete som inte skilde från vårvete (P < 0,05). Havre hade lägst TS-försvinnande båda åren. Ingen betydande effekt av utvecklingsstadium visades på TS-försvinnandet vid inkuberingen. Havre och vårvete innehöll mer in situ- och in vitro potentiellt icke nedbrytbar NDF än korn och rågvete (P < 0,01). Korn innehöll mest potentiellt nedbrytbar NDF in vitro medan det inte fanns några skillnader i innehåll av potentiellt nedbrytbar NDF mellan grödor med in situ-metoden. Havre och vårvete hade lägst innehåll av in vitro potentiellt nedbrytbar NDF (P < 0,05). Korn hade den högsta nedbrytningshastigheten av NDF in situ vid båda utvecklingsstadierna. Skillnader mellan de övriga grödorna i nedbrytningshastighet in situ var olika vid olika utvecklingsstadium (P < 0,01). Innehållet av potentiellt nedbrytbar NDF var högre vid tidig skörd än vid senare skörd både in vitro och in situ (P < 0,01). Mängden potentiellt icke nedbrytbar NDF skilde inte mellan utvecklingsstadier utom för in situ-metoden där skillnaden troligen orsakats av några felskattade värden, samt för vårvete andra året in vitro där mängden icke nedbrytbar NDF ökade från tidig mjölk- till tidig degmognad. Nedbrytningshastigheten av NDF in situ minskade med senare utvecklingsstadium (P < 0,001). Variationen i innehåll av in situ potentiellt icke nedbrytbar NDF samt TS-försvinnandet efter 120 timmars inkubering in situ kunde till ganska stor del förklaras av skillnader i ligninhalt (R2=0,49 – 0,68). Tidsfördröjningen innan nedbrytningen av NDF startade var 15 gånger längre med in vitro-metoden jämfört med in situ (P < 0,001). In vitro-metoden gav även högre nedbrytningshastighet av NDF samt lägre total nedbrytning av NDF och TS (P < 0,001). Den högre nedbrytningshastigheten med in vitro-metoden är ett resultat av den långa tidsfördröjningen innan inkuberingen började. Skillnader i nedbrytningsgrad mellan metoderna beror troligen på den högre mikrobkoncentrationen i vommen hos det levande djuret jämfört med in vitro, samt en onormalt lång tidsfördröjning på grund av brister i metodiken, såsom lång transport av vomvätska. Trots att in vitro-metoden uppvisade en större variation mellan replikat och att värden från in vitro-metoden passade sämre med smältbarhetsmodellen än in situ-metodens värden, visades en god korrelation mellan de båda metoderna vad gäller potentiellt nedbrytbar och icke nedbrytbar NDF. ANKOM DaisyII in vitro-metoden fungerade för skattning av nedbrytningsgraden av NDF i helsäd, men ytterligare utveckling av metoden behövs för att korta ner tidsfördröjningen och minska variationen, för att uppnå säkrare skattningar av nedbrytningskinetiken

Differences at brain SPECT between depressed females with and without adult ADHD and healthy controls: etiological considerations

<p>Abstract</p> <p>Background</p> <p>Comorbidity between Attention Deficit Hyperactivity Disorder (ADHD) and mood disorders is common. Alterations of the cerebellum and frontal regions have been reported in neuro-imaging studies of ADHD and major depression.</p> <p>Methods</p> <p>Thirty chronically depressed adult females of whom 16 had scores below, and 14 scores above, cut-offs on the 25-items Wender Utah Retrospective Scale (WURS-25) and the Wender-Reimherr Adult Attention Deficit Disorder Scale (WRAADDS) were divided into subgroups designated "Depression" and "Depression + ADHD", respectively. Twenty-one of the patients had some audiological symptom, tinnitus and/or hearing impairment. The patients were investigated with other rating scales and ^99mTc-HMPAO SPECT. Controls for ^99mTc-HMPAO SPECT were 16 healthy females. SPECT was analyzed by both statistical parametric mapping (SPM2) and the computerized brain atlas (CBA). Discriminant analysis was performed on the volumes of interest generated by the CBA, and on the scores from rating scales with the highest group differences.</p> <p>Results</p> <p>The mean score of a depression rating scale (MADRS-S) was significantly lower in the "Depression" subgroup compared to in the "Depression + ADHD" subgroup. There was significantly decreased tracer uptake within the bilateral cerebellum at both SPM and CBA in the "Depression + ADHD" subgroup compared to in the controls. No decrease of cerebellar tracer uptake was observed in "Depression". Significantly increased tracer uptake was found at SPM within some bilateral frontal regions (Brodmann areas 8, 9, 10, 32) in the "Depression + ADHD" subgroup compared to in "Depression". An accuracy of 100% was obtained for the discrimination between the patient groups when thalamic uptake was used in the analysis along with scores from Socialization and Impulsivity scales.</p> <p>Conclusion</p> <p>The findings confirm the previous observation of a cerebellar involvement in ADHD. Higher bilateral frontal ^99mTc-HMPAO uptake in "Depression + ADHD" compared to in "Depression" indicate a difference between these subgroups. ^99mTc-HMPAO uptake mechanisms are discussed.</p

Associations between antimicrobial susceptibility/resistance of Neisseria gonorrhoeae isolates in European Union/European Economic Area and patients' gender, sexual orientation and anatomical site of infection, 2009-2016

Background: The emergence and spread of antimicrobial resistance (AMR) in Neisseria gonorrhoeae, nationally and internationally, is a serious threat to the management and control of gonorrhoea. Limited and conflicting data regarding the epidemiological drivers of gonococcal AMR internationally have been published. We examined the antimicrobial susceptibility/resistance of gonococcal isolates (n = 15,803) collected across 27 European Union/European Economic Area (EU/EEA) countries in 2009–2016, in conjunction to epidemiological and clinical data of the corresponding patients, to elucidate associations between antimicrobial susceptibility/resistance and patients’ gender, sexual orientation and anatomical site of infection. Methods: In total, 15,803 N. gonorrhoeae isolates from the European Gonococcal Antimicrobial Surveillance Programme (Euro-GASP), 2009–2016, were examined. Associations between gonococcal susceptibility/resistance and patients’ gender, sexual orientation and anatomical site of infection were investigated using univariate and multivariate logistic regression analysis. Statistical significance was determined by Pearson χ 2-test or Fisher’s exact test with two-tailed p-values of < 0.05 indicating significance Results: The overall gonococcal resistance from 2009 to 2016 was 51.7% (range during the years: 46.5–63.5%), 7.1% (4.5–13.2%), 4.3% (1.8–8.7%), and 0.2% (0.0–0.5%) to ciprofloxacin, azithromycin, cefixime, and ceftriaxone, respectively. The level of resistance combined with decreased susceptibility to ceftriaxone was 10.2% (5.7–15.5%). Resistance to cefixime and ciprofloxacin, and resistance combined with decreased susceptibility to ceftriaxone were positively associated with urogenital infections and heterosexual males, males with sexual orientation not reported and females (except for ciprofloxacin), i.e. when compared to men-who-have-sex-with-men (MSM). Azithromycin resistance was positively associated with heterosexual males, but no association was significant regarding anatomical site of infection. Conclusions: Overall, sexual orientation was the main variable associated with gonococcal AMR. Strongest positive associations were identified with heterosexual patients, particularly males, and not MSM. To provide evidence-based understanding and mitigate gonococcal AMR emergence and spread, associations between antimicrobial susceptibility/resistance and patients’ gender, sexual orientation and anatomical site of infection need to be further investigated in different geographic settings. In general, these insights will support identification of groups at increased risk and targeted public health actions such as intensified screening, 3-site testing using molecular diagnostics, sexual contact tracing, and surveillance of treatment failures.The study was funded by the ECDC (Framework Contract No. ECDC/2017/004). Open Access funding provided by Örebro UniversityS

Evaluation of the SpeeDx ResistancePlus® GC and SpeeDx GC 23S 2611 (beta) molecular assays for prediction of antimicrobial resistance/susceptibility to ciprofloxacin and azithromycin in Neisseria gonorrhoeae

European collaborative group: Raquel Abad Torreblanca, Lena Rós Ásmundsdóttir, Eszter Balla, Irith De Baetselier, Beatrice Bercot, Thea Bergheim, Maria José Borrego, Susanne Buder, Robert Cassar, Michelle Cole, Alje van Dam, Claudia Eder, Steen Hoffmann, Blazenka Hunjak, Samo Jeverica, Vesa Kirjavainen, Panayiota Maikanti-Charalambous, Vivi Miriagou, Beata Młynarczyk-Bonikowska, Gatis Pakarna, Peter Pavlik, Monique Perrin, Joseph Pett, Paola Stefanelli, Kate Templeton, Magnus Unemo, Jelena Viktorova, Hana ZákouckáPortugal: Maria-José Borrego (INSA)Background: Accurate molecular assays for prediction of antimicrobial resistance (AMR)/susceptibility in Neisseria gonorrhoeae (Ng) can offer individualized treatment of gonorrhoea and enhanced AMR surveillance. Objectives: We evaluated the new ResistancePlus® GC assay and the GC 23S 2611 (beta) assay (SpeeDx), for prediction of resistance/susceptibility to ciprofloxacin and azithromycin, respectively. Methods: Nine hundred and sixty-seven whole-genome-sequenced Ng isolates from 20 European countries, 143 Ng-positive (37 with paired Ng isolates) and 167 Ng-negative clinical Aptima Combo 2 (AC2) samples, and 143 non-gonococcal Neisseria isolates and closely related species were examined with both SpeeDx assays. Results: The sensitivity and specificity of the ResistancePlus® GC assay to detect Ng in AC2 samples were 98.6% and 100%, respectively. ResistancePlus® GC showed 100% sensitivity and specificity for GyrA S91 WT/S91F detection and 99.8% sensitivity and specificity in predicting phenotypic ciprofloxacin resistance. The sensitivity and specificity of the GC 23S 2611 (beta) assay for Ng detection in AC2 samples were 95.8% and 100%, respectively. GC 23S 2611 (beta) showed 100% sensitivity and 99.9% specificity for 23S rRNA C2611 WT/C2611T detection and 64.3% sensitivity and 99.9% specificity for predicting phenotypic azithromycin resistance. Cross-reactions with non-gonococcal Neisseria species were observed with both assays, but the analysis software solved most cross-reactions. Conclusions: The new SpeeDx ResistancePlus® GC assay performed well in the detection of Ng and AMR determinants, especially in urogenital samples. The GC 23S 2611 (beta) assay performed relatively well, but its sensitivity, especially for predicting phenotypic azithromycin resistance, was suboptimal and further optimizations are required, including detection of additional macrolide resistance determinant(s).This work was supported by the O¨rebro County Council Research Committee and the Foundation for Medical Research at O¨rebro University Hospital, O¨rebro, Sweden.info:eu-repo/semantics/publishedVersio

WHO laboratory validation of Xpert® CT/NG and Xpert® TV on the GeneXpert system verifies high performances.

Effective tests for diagnosis of sexually transmitted infections (STIs), used point of care to inform treatment and management decisions, are urgently needed. We evaluated the analytical sensitivity and specificity of the Xpert® CT/NG and Xpert® TV tests, examining 339 samples spiked with phenotypically and/or genetically diverse strains of Neisseria gonorrhoeae, Chlamydia trachomatis, and Trichomonas vaginalis, and other related species that may cross-react. The APTIMA Combo 2 test and APTIMA TV test were used as reference tests. The analytical sensitivity for all three agents in the Xpert® CT/NG and Xpert® TV tests was ≤102 genome equivalents/reaction. The analytical specificity of both tests was high. False-positive results were identified in the Xpert® TV test when challenging with high concentrations of Trichomonas tenax, Trichomonas gallinae, Trichomonas stableri, and Trichomonas aotus. However, the clinical relevance of these cross-reactions can likely be neglected, because these species have not been identified in urogenital samples from humans. In conclusion, the analytical sensitivity and specificity of the user-friendly Xpert® CT/NG and Xpert® TV tests on the GeneXpert system were high. The results support the use of specimens from also extra-genital sites, for example, pharynx and rectum. However, appropriate clinical validations are additionally required

Associations between antimicrobial susceptibility/resistance of Neisseria gonorrhoeae isolates in European Union/European Economic Area and patients' gender, sexual orientation and anatomical site of infection, 2009-2016.

BACKGROUND: The emergence and spread of antimicrobial resistance (AMR) in Neisseria gonorrhoeae, nationally and internationally, is a serious threat to the management and control of gonorrhoea. Limited and conflicting data regarding the epidemiological drivers of gonococcal AMR internationally have been published. We examined the antimicrobial susceptibility/resistance of gonococcal isolates (n = 15,803) collected across 27 European Union/European Economic Area (EU/EEA) countries in 2009-2016, in conjunction to epidemiological and clinical data of the corresponding patients, to elucidate associations between antimicrobial susceptibility/resistance and patients' gender, sexual orientation and anatomical site of infection. METHODS: In total, 15,803 N. gonorrhoeae isolates from the European Gonococcal Antimicrobial Surveillance Programme (Euro-GASP), 2009-2016, were examined. Associations between gonococcal susceptibility/resistance and patients' gender, sexual orientation and anatomical site of infection were investigated using univariate and multivariate logistic regression analysis. Statistical significance was determined by Pearson χ2-test or Fisher's exact test with two-tailed p-values of < 0.05 indicating significance. RESULTS: The overall gonococcal resistance from 2009 to 2016 was 51.7% (range during the years: 46.5-63.5%), 7.1% (4.5-13.2%), 4.3% (1.8-8.7%), and 0.2% (0.0-0.5%) to ciprofloxacin, azithromycin, cefixime, and ceftriaxone, respectively. The level of resistance combined with decreased susceptibility to ceftriaxone was 10.2% (5.7-15.5%). Resistance to cefixime and ciprofloxacin, and resistance combined with decreased susceptibility to ceftriaxone were positively associated with urogenital infections and heterosexual males, males with sexual orientation not reported and females (except for ciprofloxacin), i.e. when compared to men-who-have-sex-with-men (MSM). Azithromycin resistance was positively associated with heterosexual males, but no association was significant regarding anatomical site of infection. CONCLUSIONS: Overall, sexual orientation was the main variable associated with gonococcal AMR. Strongest positive associations were identified with heterosexual patients, particularly males, and not MSM. To provide evidence-based understanding and mitigate gonococcal AMR emergence and spread, associations between antimicrobial susceptibility/resistance and patients' gender, sexual orientation and anatomical site of infection need to be further investigated in different geographic settings. In general, these insights will support identification of groups at increased risk and targeted public health actions such as intensified screening, 3-site testing using molecular diagnostics, sexual contact tracing, and surveillance of treatment failures

The European gonococcal antimicrobial surveillance programme (Euro-GASP) appropriately reflects the antimicrobial resistance situation for Neisseria gonorrhoeae in the European Union/European Economic Area.

BACKGROUND: European Gonococcal Antimicrobial Surveillance Programme (Euro-GASP) antimicrobial resistance (AMR) data are used to inform gonorrhoea treatment guidelines; therefore the data need to be robust and representative. We assessed the extent to which Euro-GASP reflects national measures of the AMR situation for Neisseria gonorrhoeae across the European Union/European Economic Area (EU/EEA). METHODS: We compared data from Euro-GASP with published national gonococcal AMR data from 15 countries for azithromycin, cefixime and ciprofloxacin for the period 2009 to 2013 and performed Poisson regression to identify differences (p  90%). EQA performance was also good;  4-fold from the modal MIC of the EQA isolate. CONCLUSIONS: The overall prevalence of AMR reported by Euro-GASP reflects closely the AMR situation for N. gonorrhoeae in the EU/EEA. Euro-GASP data can be used to provide robust AMR estimates to inform the European guideline for the management of gonorrhoea

Significant increase in azithromycin "resistance" and susceptibility to ceftriaxone and cefixime in Neisseria gonorrhoeae isolates in 26 European countries, 2019

Background: The European Gonococcal Antimicrobial Surveillance Programme (Euro-GASP) performs annual sentinel surveillance of Neisseria gonorrhoeae susceptibility to therapeutically relevant antimicrobials across the European Union/European Economic Area (EU/EEA). We present the Euro-GASP results from 2019 (26 countries), linked to patient epidemiological data, and compared with data from previous years. Methods: Agar dilution and minimum inhibitory concentration (MIC) gradient strip methodologies were used to determine the antimicrobial susceptibility (using EUCAST clinical breakpoints, where available) of 3239 N. gonorrhoeae isolates from 26 countries across the EU/EEA. Significance of differences compared with Euro-GASP results in previous years was analysed using Z-test and the Pearson's χ2 test was used to assess significance of odds ratios for associations between patient epidemiological data and antimicrobial resistance. Results: European N. gonorrhoeae isolates collected between 2016 and 2019 displayed shifting MIC distributions for; ceftriaxone, with highly susceptible isolates increasing over time and occasional resistant isolates each year; cefixime, with highly-susceptible isolates becoming increasingly common; azithromycin, with a shift away from lower MICs towards higher MICs above the EUCAST epidemiological cut-off (ECOFF); and ciprofloxacin which is displaying a similar shift in MICs as observed for azithromycin. In 2019, two isolates displayed ceftriaxone resistance, but both isolates had MICs below the azithromycin ECOFF. Cefixime resistance (0.8%) was associated with patient sex, with resistance higher in females compared with male heterosexuals and men-who-have-sex-with-men (MSM). The number of countries reporting isolates with azithromycin MICs above the ECOFF increased from 76.9% (20/26) in 2016 to 92.3% (24/26) in 2019. Isolates with azithromycin MICs above the ECOFF (9.0%) were associated with pharyngeal infection sites. Following multivariable analysis, ciprofloxacin resistance remained associated with isolates from MSM and heterosexual males compared with females, the absence of a concurrent chlamydial infection, pharyngeal infection sites and patients ≥ 25 years of age. Conclusions: Resistance to ceftriaxone and cefixime remained uncommon in EU/EEA countries in 2019 with a significant decrease in cefixime resistance observed between 2016 and 2019. The significant increase in azithromycin "resistance" (azithromycin MICs above the ECOFF) threatens the effectiveness of the dual therapy (ceftriaxone + azithromycin), i.e., for ceftriaxone-resistant cases, currently recommended in many countries internationally and requires close monitoring.The study was funded by the European Centre for Disease Prevention and Control (Framework Contract No. ECDC/2017/004). The funding body designed, initiated and coordinated the study as well as assisted in the interpretation of the data, development and final approval of the manuscriptS

Europe-wide expansion and eradication of multidrug-resistant Neisseria gonorrhoeae lineages: a genomic surveillance study

Background: Genomic surveillance using quality-assured whole-genome sequencing (WGS) together with epidemiological and antimicrobial resistance (AMR) data is essential to characterise the circulating Neisseria gonorrhoeae lineages and their association to patient groups (defined by demographic and epidemiological factors). In 2013, the European gonococcal population was characterised genomically for the first time. We describe the European gonococcal population in 2018 and identify emerging or vanishing lineages associated with AMR and epidemiological characteristics of patients, to elucidate recent changes in AMR and gonorrhoea epidemiology in Europe. Methods: We did WGS on 2375 gonococcal isolates from 2018 (mainly Sept 1-Nov 30) in 26 EU and EEA countries. Molecular typing and AMR determinants were extracted from quality-checked genomic data. Association analyses identified links between genomic lineages, AMR, and epidemiological data. Findings: Azithromycin-resistant N gonorrhoeae (8·0% [191/2375] in 2018) is rising in Europe due to the introduction or emergence and subsequent expansion of a novel N gonorrhoeae multi-antigen sequence typing (NG-MAST) genogroup, G12302 (132 [5·6%] of 2375; N gonorrhoeae sequence typing for antimicrobial resistance [NG-STAR] clonal complex [CC]168/63), carrying a mosaic mtrR promoter and mtrD sequence and found in 24 countries in 2018. CC63 was associated with pharyngeal infections in men who have sex with men. Susceptibility to ceftriaxone and cefixime is increasing, as the resistance-associated lineage, NG-MAST G1407 (51 [2·1%] of 2375), is progressively vanishing since 2009-10. Interpretation: Enhanced gonococcal AMR surveillance is imperative worldwide. WGS, linked to epidemiological and AMR data, is essential to elucidate the dynamics in gonorrhoea epidemiology and gonococcal populations as well as to predict AMR. When feasible, WGS should supplement the national and international AMR surveillance programmes to elucidate AMR changes over time. In the EU and EEA, increasing low-level azithromycin resistance could threaten the recommended ceftriaxone-azithromycin dual therapy, and an evidence-based clinical azithromycin resistance breakpoint is needed. Nevertheless, increasing ceftriaxone susceptibility, declining cefixime resistance, and absence of known resistance mutations for new treatments (zoliflodacin, gepotidacin) are promising.This study was supported by the European Centre for Disease Prevention and Control, the Centre for Genomic Pathogen Surveillance, the Li Ka Shing Foundation (Big Data Institute, University of Oxford), the Wellcome Genome Campus, the Foundation for Medical Research at Örebro University Hospital, and grants from Wellcome (098051 and 099202). LSB was funded by Conselleria de Sanitat Universal i Salut Pública, Generalitat Valenciana (Plan GenT CDEI-06/20-B), Valencia, Spain, and Ministry of Science, Innovation and Universities (PID2020–120113RA-I00), Spain, at the time of analysing and writing this manuscript.S