Prognostic Implications of Clinical and Imaging Diagnostic Criteria for Myocarditis.

BACKGROUND: The European Society of Cardiology (ESC), the American College of Cardiology, the American Heart Association, and expert consensus documents provide different diagnostic criteria for myocarditis. Their overlap and prognostic value have never been compared. OBJECTIVES: This study aims to assess and compare the predictive value of ESC criteria for clinically suspected myocarditis, updated Lake-Louise criteria (LLC), American Heart Association criteria for probable acute myocarditis (pAM), and expert consensus criteria for acute myocarditis (AM) and complicated myocarditis (CM). METHODS: Patients with a clinical suspicion of myocarditis referred for cardiac magnetic resonance were enrolled at 2 centers. Those with any prior cardiomyopathy were excluded. The association of composite outcome events (heart failure hospitalization, recurrent myocarditis, sustained ventricular tachycardia, or death) with ESC diagnostic criteria, LLC, pAM, AM, and CM were compared. RESULTS: Among 1,557 consecutive patients referred for cardiac magnetic resonance with possible myocarditis, 1,050 (62.6% male; 48.9 ± 16.8 years of age) were without an alternative diagnosis. Of those, 938 (89.3%) met ESC criteria for clinically suspected myocarditis, 299 (28.5%) LLC, and 356 (33.9%), 216 (20.6%), and 77 (7.3%) pAM, AM, and CM, respectively. Adverse events occurred in 161 patients (15.3%) during a median follow-up of 3.4 years. The highest annualized event rates (6.6%) were observed in patients meeting LLC, whereas negative ESC criteria indicated excellent prognosis (0.7% annualized event rate). Among all myocarditis definitions, ESC criteria and LLC were the strongest multivariable outcome predictors and had independent and incremental prognostic value (HRadjusted: 3.87; 95% CI: 1.22-12.2; P = 0.021, and HRadjusted: 2.53; 95% CI: 1.83-3.49; P < 0.001, respectively) when adjusted for clinical characteristics. CONCLUSIONS: In a real-world cohort of patients with possible myocarditis, diagnosis was reached in most patients using ESC criteria whereas only approximately one-quarter of patients reached a diagnosis with LLC. The independent prognostic value of ESC-criteria and LLC highlights the complementary role of clinical and CMR-based findings in the diagnosis and risk stratification of myocarditis.

Impact of tafamidis on myocardial function and CMR tissue characteristics in transthyretin amyloid cardiomyopathy.

AIMS: Tafamidis improves clinical outcomes in transthyretin amyloid cardiomyopathy (ATTR-CM), yet how tafamidis affects cardiac structure and function remains poorly described. This study prospectively analysed the effect of tafamidis on 12-month longitudinal changes in cardiac structure and function by cardiac magnetic resonance (CMR) compared with the natural course of disease in an untreated historic control cohort. METHODS AND RESULTS: ATTR-CM patients underwent CMR at tafamidis initiation and at 12 months. Untreated patients with serial CMRs served as reference to compare biventricular function, global longitudinal strain (GLS), LV mass and extracellular volume fraction (ECV). Thirty-six tafamidis-treated (n = 35; 97.1% male) and 15 untreated patients (n = 14; 93.3% male) with a mean age of 78.3 ± 6.5 and 76.9 ± 6.5, respectively, and comparable baseline characteristics were included. Tafamidis was associated with preserving biventricular function (LVEF (%): 50.5 ± 12 to 50.7 ± 11.5, P = 0.87; RVEF (%): 48.2 ± 10.4 to 48.2 ± 9.4, P = 0.99) and LV-GLS (-9.6 ± 3.2 to -9.9 ± 2.4%; P = 0.595) at 12 months, while a significantly reduced RV-function (50.8 ± 7.3 to 44.2 ± 11.6%, P = 0.028; P (change over time between groups) = 0.032) and numerically worsening LVGLS (-10.9 ± 3.3 to -9.1 ± 2.9%, P = 0.097; P (change over time between groups) = 0.048) was observed without treatment. LV mass significantly declined with tafamidis (184.7 ± 47.7 to 176.5 ± 44.3 g; P = 0.011), yet remained unchanged in untreated patients (163.8 ± 47.5 to 171.2 ± 39.7 g P = 0.356, P (change over time between groups) = 0.027). Irrespective of tafamidis, ECV and native T1-mapping did not change significantly from baseline to 12-month follow-up (P > 0.05). CONCLUSIONS: Compared with untreated ATTR-CM patients, initiation of tafamidis preserved CMR-measured biventricular function and reduced LV mass at 12 months. ECV and native T1-mapping did not change significantly comparable to baseline in both groups.

Example for process validation in biobanking: Fit for purpose testing of a cryopreservation method without isopentane.

Background: The freezing process of tissue samples is crucial for the preservation of morphological and molecular features. Several biobanking guidelines describe freezing techniques for optimal outcomes. As the Vetbiobank standard freezing protocol does not comply with those recommendations in detail, a process validation was performed to demonstrate that samples are suitable for downstream applications. Here we give a formal example of a process validation in the biobanking setting, as required by the biobanking guideline ISO 20387 (2018). Methods: Three different freezing protocols, freezing in liquid nitrogen, freezing via isopentane precooled on dry ice and freezing via liquid nitrogen vapor, were assessed based on morphological integrity of mouse liver and muscle tissue samples. Samples were either frozen in cryotubes (without Optimal Cutting Temperature compound, OCT) or in cryomolds (with OCT). The protocol providing the best results was validated for reproducibility and robustness in terms of defined acceptance criteria for morphological evaluability, A260/A280 ratio, and RNA integrity number values (RIN). In addition, performance tests were run by gene expression analyzes of selected, tissue specific biomarkers to confirm that processed samples are fit for purpose. Results: From the three applied freezing protocols, freezing in liquid nitrogen generated best results. Reproducibility acceptance criteria were met for both, morphological integrity and RNA quality. The freezing method was robust for the tested tissue types and the application of OCT, with exception of liver tissue, where it led to a significant decrease of the RIN value. Gene expression analyzes showed good comparability of results regardless of the applied freezing method. Conclusion: Freezing of tissue samples in liquid nitrogen provides samples of adequate quality for subsequent RNA investigations. A negative impact of OCT on the RIN value of liver samples was observed, which was independent from the applied freezing protocol and showed no impact on subsequent gene expression analysis.

Third-generation continuous-flow left ventricular assist devices: a comparative outcome analysis by device type.

AIMS: Continuous-flow left ventricular assist devices (CF-LVADs) have become a standard of care in end-stage heart failure. Limited data exist comparing outcomes of HeartMate3 (HM3) and HeartWare HVAD (HW). We aimed to compare midterm outcomes of these devices. METHODS AND RESULTS: Investigator-initiated retrospective-observational comparative analysis of all patients who underwent primary LVAD implantation of either HM3 or HW at our centre between January 2010 and December 2020. Data were derived from a prospective registry. Primary endpoints were all-cause mortality and heart transplantation. Secondary endpoints included device-related major adverse cardiac and cerebrovascular events, which included major bleeding, major neurological dysfunction (defined as persisting neurological impairment for ≥24 h), device-related major infection (excluding driveline infections), major device malfunctions leading to re-intervention or partial device exchange (pump failure, outflow-graft twist or failure, controller failure, battery failure, patient cable failure, but excluding pump thrombosis), and pump thrombosis. Further secondary endpoints included right heart failure, gastrointestinal bleeding, driveline infections, and surgical re-interventions. The secondary outcomes were analysed not only for the first event but also for recurrent events. The analysis included competing risks analysis and recurrent event regression analysis, with adjustment for confounders age, gender, body mass index (BMI), and Interagency Registry for Mechanically Assisted Circulatory Support (INTERMACS) level. Out of 106 primary CF-LVAD implantations, 36 (34%) received HM3 and 70 (66%) received HW. Median follow-up was 1.48 years [interquartile range 0.67, 2.41]. HM3 was more often implanted in men (91.7% vs. 72.9%, P = 0.024); patients were older (median 61 years [54, 66.5] vs. 52.5 years [43, 60], P < 0.001), had a higher BMI (median 26.7 kg/m2 [23.4, 29.0] vs. 24.3 kg/m2 [20.7, 27.4], P = 0.013), had more comorbidities, and were more likely targeted for destination therapy (36.1% vs. 14.3%, P = 0.010). Death occurred in 33.3% of HM3 patients, compared with 22.9% of HW patients, P = 0.247 (probability of survival at 4 years, 54.7% vs. 74.1%, P = 0.296). After adjustment for confounders, we observed a significant six-fold risk increase in device malfunctions for HW [hazard ratio (HR) 6.49, 95% confidence interval (CI) [1.89, 22.32], P = 0.003], but no significant differences in pump thrombosis (P = 0.173) or overall survival (P = 0.801). CONCLUSIONS: Comparing midterm outcomes between HM3 and HW for LVAD support from a prospective registry, HW patients had a significantly higher risk of device malfunctions. No significant differences were evident between devices in overall survival and in respect to most outcomes.

Pump thrombosis and dynamic outflow graft compression: complications in left ventricular assist device therapy.

Over the past decade, left ventricular assist device (VAD) therapy has become more prevalent and increasingly safe. Severe complications, such as VAD pump thrombosis and outflow graft obstruction, are rare, yet still associated with high morbidity and mortality. Clinical presentation, VAD alarm and log files, laboratory analysis, and non-invasive cardiac imaging are crucial for establishing the correct diagnosis and determining clinical management. Early intervention is critical to prevent adverse cardiac remodelling or VAD pump failure.

Is it safe to irradiate the newest generation of ventricular assist devices? A case report and systematic literature review.

An increasing number of mechanical assist devices, especially left ventricular assist devices (VADs), are being implanted for prolonged periods and as destination therapy. Some VAD patients require radiotherapy due to concomitant oncologic morbidities, including thoracic malignancies. This raises the potential of VAD malfunction via radiation-induced damage. So far, only case reports and small case series on radiotherapy have been published, most of them on HeartMate II (HMII, Abbott, North Chicago, IL, USA). Significantly, the effects of irradiation on the HeartMate 3 (HM3, Abbott) remain undefined, despite the presence of controller components engineered within the pump itself. We report the first case of a patient with a HM3 who successfully underwent stereotactic hypofractionated radiotherapy due to an early-stage non-small-cell lung cancer. The patient did not suffer from any complications, including toxicity or VAD malfunction. Based on this case report and on published literature, we think that performing radiotherapy after VAD implantation with the aid of a multidisciplinary team could be performed, but more in vitro studies and cases series are needed to reinforce this statement.

[Pharmacological therapy of heart failure with reduced ejection fraction]. / Medikamentöse Therapie der Herzinsuffizienz mit verminderter Auswurffraktion.

Pharmacological therapy of heart failure with reduced ejection fraction Abstract. Pharmacological therapy for heart failure has made great progress over the last three decades and evidence-based therapies have significantly improved survival and quality of life. Angiotensin-converting enzyme inhibitors or angiotensin receptor blockers and beta-blockers are the cornerstone of the heart failure therapy; indicated in virtually every patient with heart failure and reduced ejection fraction. As soon as the left ventricular ejection fraction decreases below 35 % and / or symptoms are still present (NYHA II-IV), a mineralocorticoid receptor antagonist should be added. A rather recent addition to current heart failure therapy with convincing data is the substance combination sacubitril / valsartan. It is indicated for patients with persistent symptomatic heart failure despite optimal medical therapy with ACE inhibitors or ARBs, beta-blockers, and MRAs. Crucial for all mentioned substances is to aim for the maximal tolerated dose. Various additional therapies have no proven survival benefit but are important for symptom control in everyday life. Above all the diuretics, where loop diuretics show a better effect profile compared to thiazide diuretics. Furthermore, achieving an optimal iron status (the limit to start a substitution is significantly higher than in patients without heart failure), decreasing the heart frequency with Ivabradine (if heart rate persists above 70 / min despite fully dosed betablocker) and «lifestyle changes¼ can add to the success of the medical treatment. The importance of digoxin has been steadily decreasing. The previously advocated therapeutic anticoagulation in patients with severely reduced LVEF is not propagated anymore. Significant arrhythmias (especially atrial fibrillation and ventricular arrhythmias) are common in advanced diseases. In addition to beta-blockers, amiodarone is clearly the antiarrhythmic drug of choice. According to latest data, an early interventional treatment of atrial fibrillation by pulmonary vein ablation may be beneficial and has the potential to reduce mortality in special subgroups of patients. New developments in the field of antidiabetic drugs seem to be promising for reduction of mortality and hospitalization in patients with heart failure.

The Potential Impact of Functional Imaging on Decision Making and Outcome in Patients Undergoing Surgical Revascularization.

OBJECTIVE: Coronary angiography (CA) remains the standard for preoperative planning for surgical revascularization. However, besides anatomical imaging, current guidelines recommend additional functional imaging before a therapy decision is made. We assess the impact of functional imaging on the strategy of coronary artery bypass grafting (CABG) with particular regards on postoperative patency and myocardial perfusion. METHODS: After CA, 55 patients (47 males/8 females; age: 65.1 ± 9.5 years) underwent perfusion cardiovascular magnetic resonance (CMR) and dual-source computed tomography (DSCT) before isolated CABG (n = 31), CABG and concomitant valve surgery (valve + CABG; n = 10) and isolated valve surgery (n = 14; control). DSCT was used for analysis of significant stenosis, CMR for myocardial-perfusion to discriminate between: no ischemia (normal), ischemia, or scar. The results, unknown to the surgeons, were compared with CA and related to the location and number of distal anastomoses. Nineteen CABG patients underwent follow-up CMR and DSCT (FU: 13 ± 3 months) to compare the preop findings with the postop outcomes. RESULTS: Thirty-nine patients either received CABG alone (n = 31) or a combined procedure (n = 10) with a total of 116 distal anastomoses. DSCT was compared with CA regarding accuracy of coronary stenosis and showed 91% sensitivity, 88% specificity, and negative/positive predictive values of 89/90%. In total, 880 myocardial segments (n = 55, 16 segments/patient) were assessed by CMR. In 17% (149/880) of segments ischemia and in 8% (74/880) scar tissue was found. Interestingly, 14% (16/116) of bypass-anastomoses were placed on non-ischemic myocardium and 3% (4/116) on scar tissue. In a subgroup of 19 patients 304 segments were evaluated. Thirty-nine percent (88/304) of all segments showed ischemia preoperatively, while 94% (83/88) of these ischemic segments did not show any ischemia postoperatively. In regard to performed anastomoses, 79% of all grafts (49/62) were optimally placed, whereas 21% (13/62) were either placed into non-ischemic myocardium or scar tissue, including 10% occluded grafts (6/62). CONCLUSION: In the whole cohort analysis, 17% of grafts were placed in regions with either no ischemia or scar tissue. The subgroup analysis revealed that 94% of all ischemic segments were successfully revascularized after CABG. Thus, functional imaging could be a promising tool in preoperative planning of revascularization strategy. Avoidance of extensive and unnecessary grafting could further optimize outcomes after CABG.

Adenosine stress high-pitch 128-slice dual-source myocardial computed tomography perfusion for imaging of reversible myocardial ischemia: comparison with magnetic resonance imaging.

BACKGROUND: Coronary computed tomography angiography (CTA) enables accurate anatomic evaluation of coronary artery stenosis but lacks information about hemodynamic significance. The aim of this study was to evaluate 128-slice myocardial CT perfusion (CTP) imaging with adenosine stress using a high-pitch mode, in comparison with cardiac MRI (CMR). METHODS AND RESULTS: Thirty-nine patients with intermediate to high coronary risk profile underwent adenosine stress 128-slice dual source CTP (128×0.6 mm, 0.28 seconds). Among those, 30 patients (64 ± 10 years, 6% women) also underwent adenosine stress CMR (1.5T). The 2-step CTP protocol consisted of (1) adenosine stress-CTP using a high-pitch factor (3.4) ECG-synchronized spiral mode and (2) rest-CTP/coronary-CTA using either high-pitch (heart rate <63 bpm) or prospective ECG-triggering (heart rate >63 bpm). Results were compared with CMR and with invasive angiography in 25 patients. The performance of stress-CTP for detection of myocardial perfusion defects compared with CMR was sensitivity, 96%; specificity, 88%; positive predictive value (PPV), 93%; negative predictive value (NPV), 94% (per vessel); and sensitivity, 78%; specificity, 87%; PPV, 83%; NPV, 84% (per segment). The accuracy of stress-CTP for imaging of reversible ischemia compared with CMR was sensitivity, 95%; specificity, 96%; PPV, 95%; and NPV, 96% (per vessel). In 25 patients who underwent invasive angiography, the accuracy of CTA for detection of stenosis >70% was (per segment): sensitivity, 96%; specificity, 88%; PPV, 67%; and NPV, 98.9%. The accuracy improved from 84% to 95% after adding stress CTP to CTA. Radiation exposure of the entire stress/rest CT protocol was only 2.5 mSv. CONCLUSIONS: Adenosine-induced stress 128-slice dual-source high-pitch myocardial CTP allows for simultaneously assessment of reversible myocardial ischemia and coronary stenosis, with good diagnostic accuracy as compared with CMR and invasive angiography, at a very low radiation exposure.

Delayed enhancement imaging of myocardial viability: low-dose high-pitch CT versus MRI.

OBJECTIVES: To evaluate the accuracy of high-pitch delayed enhancement (DE) CT for the assessment of myocardial viability with MRI as the reference standard. METHODS: Twenty-four patients (mean age 66.9 ± 9.2 years) with coronary artery disease underwent DE imaging with 128-slice dual-source CT (prospective electrocardiography (ECG)-triggering) and MRI at 1.5 T. Two observers assessed DE transmurality per segment, and measured signal intensity (MRI) or attenuation (CT) in infarcted and healthy myocardium and noise in the left ventricular blood pool for calculating contrast-to-noise ratios (CNR). RESULTS: 75/408 (18.4%) segments in 18/24 patients (75.0%) showed DE in MRI, of which 28 segments in 10/24 (41.7%) patients were non-viable (scar tissue transmurality >50%). Sensitivity, specificity and accuracy of CT for diagnosis of non-viability were 60.7%, 96.8% and 94.4% per segment, and 90.0%, 92.9% and 91.7% per patient. CNR was significantly higher in MR (7.4 ± 3.0 vs. 4.6 ± 1.5; p = 0.018), and image noise significantly lower (11.6 ± 5.7 vs.15.0 ± 4.5; p = 0.019). Radiation dose of DECT was 0.89 ± 0.07 mSv. CONCLUSIONS: CTDE imaging in the high-pitch mode enables myocardial viability assessment at a low radiation dose and good accuracy compared with MR, although associated with a lower CNR and higher noise.

Dual-step prospective ECG-triggered 128-slice dual-source CT for evaluation of coronary arteries and cardiac function without heart rate control: a technical note.

PURPOSE: To describe prospective ECG-triggered dual-source CT dual-step pulsing (pECG(dual_step)) for evaluation of coronary arteries and cardiac function. METHODS: Fifty-one consecutive patients pre- or post-cardiovascular surgery were examined with adaptive sequential tube current modulated (pECG(dual-step)) 128-slice dual-source CT without heart rate control (main padding window: 40% RR interval >65 bpm/70% RR interval <65 bpm). Image quality of coronary arteries was graded (4-point scale), and cardiac function was evaluated. RESULTS: Mean HR was 68 bpm. Thirty-seven patients were in stable sinus rhythm (SR); 14 had arrhythmia. Image quality of coronary arteries was diagnostic in 804/816 (98%) of segments. The number of non-diagnostic segments was higher in patients with arrhythmia as compared to those in SR (4% vs. 0.5%; p = 0.01), and there were fewer segments with excellent image quality (79% vs. 94%; p < 0.001) and more segments with impaired image quality (p < 0.001 and p = 0.002). Global and regional LV function could be evaluated in 41 (80%) and 47 (92%) patients, and valvular function in 48 (94%). In 11/14 of patients with arrhythmia, the second step switched to full mAs, increasing radiation exposure to 8.6 mAs (p < 0.001). The average radiation dose was 3.8 mSv (range, 1.7-7.9) in patients in SR. CONCLUSION: pECG(dual-step)128-slice DSCT is feasible for the evaluation of coronary arteries and cardiac function without heart rate control in patients in stable sinus rhythm at a low radiation dose.

Low dose high-pitch spiral acquisition 128-slice dual-source computed tomography for the evaluation of coronary artery bypass graft patency.

OBJECTIVES: To prospectively assess electrocardiography (ECG)-synchronized dual-source computed tomography (CT) in high-pitch spiral acquisition mode for the evaluation of coronary artery bypass graft (CABG) patency regarding image quality and radiation dose. MATERIALS AND METHODS: Fifty consecutive patients (47 men, age 69.6 +/- 9.6 years, body mass index 26.6 +/- 3.5 kg/m) underwent clinically indicated 128-slice dual-source CT angiography of the entire thorax for the evaluation of graft patency after CABG surgery using a prospectively ECG-synchronized high-pitch spiral acquisition mode (pitch 3.2; 100 kV tube voltage, 0.28 seconds gantry rotation time). Heart rates (HR) were noted. Image quality of the proximal anastomosis, graft body, distal anastomosis, and postanastomotic coronary artery were graded by 2 independent readers on a 3-point scale (1 = excellent, 2 = moderate, and 3 = poor/nondiagnostic). Quantitative image quality parameters (noise, contrast-to-noise ratio) were measured. Effective radiation doses were calculated. RESULTS: Mean HR was 76 +/- 19 bpm (range, 45-135 bpm). Median scan time was 0.9 seconds (range, 0.76-1.17 seconds), mean scan length was 349 +/- 38 mm (range, 294-452 mm). A total of 125 CABG (54% arterial grafts) and 465 vessel sections were analyzed. Twelve grafts were proximally occluded. Diagnostic image quality was obtained in 462 (99.4%) sections. Of those, image quality was excellent in 397 (85.4%) and moderate in 65 (14.0%). All 3 (0.6%) nondiagnostic sections affected the distal anastomosis due to motion artifacts caused by high HR. Separating the study population by the median HR of 71 bpm, image quality of the distal anastomosis regarding motion artifacts was significantly lower at high HR (P < 0.05). Interobserver agreement in grading image quality of graft segments was good (kappa = 0.77). Image noise in the aorta was 36.9 +/- 8.1, contrast-to-noise ratio was 13.1 +/- 4.2. Effective radiation dose was 2.3 +/- 0.3 mSv. CONCLUSIONS: The patency of coronary artery bypass grafts can be assessed with decreasing image quality at high HR in high-pitch prospectively ECG-synchronized thoracic 128-slice dual-source CT angiography at a low radiation dose.

Sphingomonas fennica sp. nov. and Sphingomonas haloaromaticamans sp. nov., outliers of the genus Sphingomonas.

Bacterial isolates obtained from polychlorophenol-contaminated sites in Finland (strain K101(T)) and from a Dutch drinking water well (strain A175(T)) were characterized taxonomically. 16S rRNA gene sequence analysis, determination of DNA G+C content, physiological characterization, estimation of the ubiquinone and polar lipid patterns and fatty acid content revealed that strains K101(T) and A175(T) were similar to Sphingomonas wittichii RW1(T) but also showed pronounced differences. The DNA G+C contents of the two novel strains were 63.6 and 66.1 mol%, respectively. On the basis of these results, two novel species of the genus Sphingomonas are described, for which the names Sphingomonas haloaromaticamans sp. nov. [type strain A175(T) (=DSM 13477(T)=CCUG 53463(T))] and Sphingomonas fennica sp. nov. [type strain K101(T) (=DSM 13665(T)=CCUG 53462(T))] are proposed.

Sphingobium aromaticiconvertens sp. nov., a xenobiotic-compound-degrading bacterium from polluted river sediment.

A bacterial strain capable of degrading some monochlorinated dibenzofurans, designated RW16T, was isolated from aerobic River Elbe sediments. The strain was characterized based on 16S rRNA gene sequence analysis, DNA G+C content, physiological characteristics, polyamines, ubiquinone and polar lipid pattern and fatty acid composition. This analysis revealed that strain RW16T represents a novel species of the genus Sphingobium. The DNA G+C content of strain RW16T, 60.7 mol%, is the lowest yet reported for the genus. 16S rRNA gene sequence analysis placed strain RW16T as an outlier in the genus Sphingobium. The name Sphingobium aromaticiconvertens sp. nov. is proposed for this dibenzofuran-mineralizing organism, with type strain RW16T (=DSM 12677T=CIP 109198T).

Bacillus herbersteinensis sp. nov.

Two bacterial strains, designated D-1,5a(T) and D-1,5b, were isolated from a medieval wall painting in the chapel of Castle Herberstein, Styria (Austria). The Gram-positive, heterotrophic, aerobic, spore-forming rods showed nearly identical whole-cell protein patterns, identical genomic fingerprints and identical physiological profiles, demonstrating their relationship at the species level. Both strains contained meso-diaminopimelic acid in their peptidoglycan, possessed a quinone system comprising menaquinone MK-7 and had fatty acid profiles in which C(15:0) iso and C(15:0) anteiso were predominant. The 16S rRNA gene sequence of D-1,5a(T) showed the highest similarity (99.5%) to the sequence of Bacillus sp. LMG 20243, and Bacillus flexus IFO 15715(T) was the next most closely related established species (96.5%). Other type strains, such as Bacillus fastidiosus DSM 91(T), Bacillus indicus SD/3(T), Bacillus cibi JG-30(T), Bacillus megaterium IAM 13418(T), Bacillus cohnii DSM 6308(T), Bacillus bataviensis LMG 21833(T) and Bacillus soli LMG 21838(T), shared 96.0-96.1% 16S rRNA gene sequence similarity with D-1,5a(T). The combination of physiological and chemotaxonomic traits distinguishes the two strains from those species sharing the highest sequence similarities (96.0-96.5%). On the basis of these characteristics and the phylogenetic position of strain D-1,5a(T) (=DSM 16534(T)=CCM 7228(T)), this strain is assigned as the type strain of a novel species of the genus Bacillus, for which the name Bacillus herbersteinensis sp. nov. is proposed.

Staphylococcus nepalensis sp. nov., isolated from goats of the Himalayan region.

Four coagulase-negative, novobiocin-resistant cocci, designated CW1(T), PM34, MM3 and RW78, were isolated from the respiratory tract of goats kept in the Himalayan region. The four isolates were assigned to a single species on the basis of almost identical biochemical and physiological traits, protein profiles obtained after SDS-PAGE and identical genomic fingerprints generated after enterobacterial repetitive intergenic consensus (ERIC)-PCR. Strain CW1(T) showed highest 16S rDNA sequence similarities to Staphylococcus cohnii subsp. urealyticus ATCC 49330(T), Staphylococcus saprophyticus subsp. saprophyticus ATCC 15305(T), S. cohnii subsp. cohnii ATCC 29974(T), Staphylococcus arlettae ATCC 43957(T), Staphylococcus gallinarum ATCC 35539(T), Staphylococcus succinus ATCC 700337(T) and Staphylococcus xylosus ATCC 29971(T) (99.0, 98.8, 98.8, 98.4, 98.2, 98.1 and 98.1 %, respectively), indicating its classification within the genus Staphylococcus. The polar lipid composition, fatty acid profiles, quinone systems and diagnostic cell-wall diamino acid were in agreement with the characteristics of the genus Staphylococcus. DNA-DNA hybridization with closely related Staphylococcus species suggested that strain CW1(T) represents an as-yet unrecognized species. Based on these results, a novel species of the genus Staphylococcus is described, Staphylococcus nepalensis sp. nov. The type strain is CW1(T) (=DSM 15150(T)=CCM 7045(T)) and the most dissimilar strain is PM34 (=DSM 15151=CCM 7046).