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BACKGROUND: Multiple sclerosis (MS) negatively affects health-related quality of life (HRQoL). OBJECTIVE: To evaluate HRQoL in people with highly active relapsing MS treated with cladribine tablets (CladT; 3.5 mg/kg cumulative dose over 2 years) in CLARIFY-MS. METHODS: Changes in the MS quality of life (MSQoL)-54 scores were analysed using a repeated mixed-effects linear model. Subgroup analyses were performed for participants who were pretreatment-naïve and those pretreated with disease-modifying therapies (DMTs) before initiating CladT. Safety and tolerability of CladT were also assessed. RESULTS: MSQoL-54 physical (mean change = 4.86; 95% confidence interval (CI) = 3.18, 6.53) and mental health (4.80; 95% CI = 3.13, 6.46) composite scores (primary endpoints) showed significant improvement at Month 24 versus Baseline (p < 0.0001). Changes in the MSQoL-54 scores were consistent across the pretreatment-naïve and DMT-pretreated subgroups. No new severe or opportunistic infections occurred. Most post-baseline lymphopenia events were Grade 1-2 in severity. Transient Grade-3 lymphopenia was observed in 19.7% (95/482) of participants. Grade-4 lymphopenia was not observed. CONCLUSIONS: CladT treatment significantly improved the mean MSQoL-54 physical and mental health composite scores over 2 years. CladT efficacy in HRQoL, relapse rates and Expanded Disability Status Scale scores demonstrates its multidimensional effects in MS treatment.
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Linfopenia , Esclerosis Múltiple Recurrente-Remitente , Esclerosis Múltiple , Humanos , Cladribina/efectos adversos , Esclerosis Múltiple/tratamiento farmacológico , Inmunosupresores/efectos adversos , Calidad de Vida , Esclerosis Múltiple Recurrente-Remitente/tratamiento farmacológico , Linfopenia/inducido químicamente , Linfopenia/tratamiento farmacológico , Comprimidos/uso terapéuticoRESUMEN
BACKGROUND: In the CLARITY (CLAdRIbine Tablets treating multiple sclerosis orallY) study of patients with relapsing-remitting multiple sclerosis, treatment with cladribine tablets 3.5 mg/kg (CladT) significantly reduced the annualised relapse rate (ARR) versus placebo; this effect was sustained in CLARITY Extension, without further treatment. OBJECTIVE: To assess the frequency and severity of relapses in patients treated with CladT versus placebo in CLARITY over 2 years and evaluate the durability of effect in patients who received no further treatment for 2 years in CLARITY Extension. METHODS: In this post hoc analysis, ARRs were calculated for qualifying and all relapses, and qualifying and all severe relapses (i.e. requiring steroid treatment or leading to hospitalisation) in patients treated with CladT (n = 433) and placebo (n = 437) in CLARITY, and those from the CladT group who received placebo in CLARITY Extension (n = 98). RESULTS: At Month 6, Year 1 and Year 2, patients receiving CladT had a significantly lower risk of qualifying or all relapses (all p < 0.0001), and qualifying or all severe relapses (all p < 0.005), compared with placebo. This effect was sustained in CLARITY Extension without further treatment. CONCLUSION: The results show durable efficacy of cladribine tablets 3.5 mg/kg for reducing frequency and severity of relapses in patients with relapsing-remitting multiple sclerosis.CLARITY: NCT00213135; CLARITY Extension: NCT00641537.
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Esclerosis Múltiple Recurrente-Remitente , Esclerosis Múltiple , Cladribina/uso terapéutico , Humanos , Inmunosupresores/uso terapéutico , Esclerosis Múltiple/tratamiento farmacológico , Esclerosis Múltiple Recurrente-Remitente/tratamiento farmacológico , Recurrencia , ComprimidosRESUMEN
OBJECTIVE: We assessed the efficacy and safety of once-daily eslicarbazepine acetate in comparison with twice-daily (BID) controlled-release carbamazepine (carbamazepine-CR) monotherapy in newly diagnosed focal epilepsy patients. METHODS: This randomized, double-blind, noninferiority trial (NCT01162460) utilized a stepwise design with 3 dose levels. Patients who remained seizure-free for the 26-week evaluation period (level A: eslicarbazepine acetate 800 mg/carbamazepine-CR 200 mg BID) entered a 6-month maintenance period. If a seizure occurred during the evaluation period, patients were titrated to the next target level (level B: eslicarbazepine acetate 1200 mg/carbamazepine-CR 400 mg BID, level C: eslicarbazepine acetate 1600 mg/carbamazepine-CR 600 mg BID) and the evaluation period began again. The primary endpoint was the proportion of seizure-free patients for 6 months after stabilization in the per protocol set. The predefined noninferiority criteria were -12% absolute and -20% relative difference between treatment groups. RESULTS: Eight hundred fifteen patients were randomly assigned; 785 (388 in the eslicarbazepine acetate group and 397 in the carbamazepine-CR group) were included in the per protocol set, and 813 (401 in the eslicarbazepine acetate group and 412 in the carbamazepine-CR group) were included in the full analysis set for the primary analysis. Overall, 71.1% of eslicarbazepine acetate-treated patients and 75.6% of carbamazepine-CR-treated patients were seizure-free for ≥6 months at the last evaluated dose (average risk difference = -4.28%, 95% confidence interval [CI] = -10.30 to 1.74; relative risk difference = -5.87%, 95% CI = -13.50 to 2.44) in the per protocol set. Rates of treatment-emergent adverse events were similar between groups for patients in the safety set. Noninferiority was also demonstrated in the full analysis set, as 70.8% of patients with eslicarbazepine acetate and 74.0% with carbamazepine-CR were seizure-free at the last evaluated dose (average risk difference = -3.07, 95% CI = -9.04 to 2.89). SIGNIFICANCE: Treatment with eslicarbazepine acetate was noninferior to BID carbamazepine-CR. With its once-daily formulation, eslicarbazepine acetate provides a useful option for first-line monotherapy for adults with newly diagnosed epilepsy and focal onset seizures.
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Anticonvulsivantes/administración & dosificación , Carbamazepina/administración & dosificación , Dibenzazepinas/administración & dosificación , Epilepsias Parciales/tratamiento farmacológico , Bloqueadores del Canal de Sodio Activado por Voltaje/administración & dosificación , Adulto , Alanina Transaminasa/sangre , Anticonvulsivantes/uso terapéutico , Carbamazepina/uso terapéutico , Preparaciones de Acción Retardada , Dibenzazepinas/uso terapéutico , Mareo/inducido químicamente , Relación Dosis-Respuesta a Droga , Método Doble Ciego , Estudios de Equivalencia como Asunto , Fatiga/inducido químicamente , Femenino , Cefalea/inducido químicamente , Humanos , Modelos Logísticos , Masculino , Persona de Mediana Edad , Náusea/inducido químicamente , Resultado del Tratamiento , Bloqueadores del Canal de Sodio Activado por Voltaje/uso terapéutico , Adulto Joven , gamma-Glutamiltransferasa/sangreRESUMEN
Injection of Yersinia outer proteins (Yops) into host cells by a type III secretion system is an important immune evasion mechanism of Yersinia enterocolitica (Ye). In this process Ye invasin (Inv) binds directly while Yersinia adhesin A (YadA) binds indirectly via extracellular matrix (ECM) proteins to ß1 integrins on host cells. Although leukocytes turned out to be an important target of Yop injection by Ye, it was unclear which Ye adhesins and which leukocyte receptors are required for Yop injection. To explain this, we investigated the role of YadA, Inv and ß1 integrins for Yop injection into leukocytes and their impact on the course of systemic Ye infection in mice. Ex vivo infection experiments revealed that adhesion of Ye via Inv or YadA is sufficient to promote Yop injection into leukocytes as revealed by a ß-lactamase reporter assay. Serum factors inhibit YadA- but not Inv-mediated Yop injection into B and T cells, shifting YadA-mediated Yop injection in the direction of neutrophils and other myeloid cells. Systemic Ye mouse infection experiments demonstrated that YadA is essential for Ye virulence and Yop injection into leukocytes, while Inv is dispensable for virulence and plays only a transient and minor role for Yop injection in the early phase of infection. Ye infection of mice with ß1 integrin-depleted leukocytes demonstrated that ß1 integrins are dispensable for YadA-mediated Yop injection into leukocytes, but contribute to Inv-mediated Yop injection. Despite reduced Yop injection into leukocytes, ß1 integrin-deficient mice exhibited an increased susceptibility for Ye infection, suggesting an important role of ß1 integrins in immune defense against Ye. This study demonstrates that Yop injection into leukocytes by Ye is largely mediated by YadA exploiting, as yet unknown, leukocyte receptors.
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Adhesinas Bacterianas/fisiología , Proteínas de la Membrana Bacteriana Externa/administración & dosificación , Integrina beta1/fisiología , Leucocitos/metabolismo , Yersiniosis/sangre , Yersinia enterocolitica , Adhesinas Bacterianas/genética , Alelos , Animales , Integrina beta1/genética , Ratones , Ratones Endogámicos C57BL , PlásmidosRESUMEN
The current paradigm suggests that Yersinia enterocolitica (Ye) adheres to host cells via the outer membrane proteins Yersinia adhesin A (YadA) or invasin (Inv) to facilitate injection of Yops by the type III secretion system. In this process Inv binds directly to ß1 integrins of host cells while YadA may bind indirectly via extracellular matrix proteins to ß1 integrins. Here we challenged this paradigm and investigated the requirements for Yop injection. We demonstrate that Inv- but not YadA-mediated adhesion depends on ß1 integrin binding and activation, and that tight adhesion is a prerequisite for Yop injection. By means of novel transgenic cell lines, shRNA approaches and RGD peptides, we found that YadA, in contrast to Inv, may use a broad host cell receptor repertoire for host cell adhesion. In the absence of ß1 integrins, YadA mediates Yop injection by interaction with αV integrins in cooperation with yet unknown cofactors expressed by epithelial cells, but not fibroblasts. Electron microscopic and flow chamber studies revealed that a defined intimate contact area between Ye and host cells resulting in adhesion forces resisting shear stress is required for Yop injection. Thus, the indirect binding of YadA to a broad extracellular matrix (ECM) binding host cell receptor repertoire of different cell types makes YadA a versatile tool to ensure Yop injection. In conclusion, given the differential expression of the outer membrane proteins Inv and YadA in the course of Ye infection and differential expression of integrins by various host cell populations, the data demonstrate that Ye is flexibly armed to accomplish Yop injection in different host cell types, a central event in its immune evasion strategy.
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Adhesinas Bacterianas/metabolismo , Adhesión Bacteriana , Toxinas Bacterianas/metabolismo , Interacciones Huésped-Patógeno , Yersinia enterocolitica/fisiología , Células Epiteliales/microbiología , Fibroblastos/metabolismo , Citometría de Flujo , Integrina alfaV/metabolismo , Integrina beta1/metabolismo , Microscopía Electrónica , Unión Proteica , Transporte de ProteínasRESUMEN
CD4(+) T cells are essential for the control of Yersinia enterocolitica (Ye) infection in mice. Ye can inhibit dendritic cell (DC) antigen uptake and degradation, maturation and subsequently T-cell activation in vitro. Here we investigated the effects of Ye infection on splenic DCs and T-cell proliferation in an experimental mouse infection model. We found that OVA-specific CD4(+) T cells had a reduced potential to proliferate when stimulated with OVA after infection with Ye compared to control mice. Additionally, proliferation of OVA-specific CD4(+) T cells was markedly reduced when cultured with splenic CD8α(+) DCs from Ye infected mice in the presence of OVA. In contrast, T-cell proliferation was not impaired in cultures with CD4(+) or CD4(-)CD8α(-) DCs isolated from Ye infected mice. However, OVA uptake and degradation as well as cytokine production were impaired in CD8α(+) DCs, but not in CD4(+) and CD4(-)CD8α(-) DCs after Ye infection. Pathogenicity factors (Yops) from Ye were most frequently injected into CD8α(+) DCs, resulting in less MHC class II and CD86 expression than on non-injected CD8α(+) DCs. Three days post infection with Ye the number of splenic CD8α(+) and CD4(+) DCs was reduced by 50% and 90%, respectively. The decreased number of DC subsets, which was dependent on TLR4 and TRIF signaling, was the result of a faster proliferation and suppressed de novo DC generation. Together, we show that Ye infection negatively regulates the stimulatory capacity of some but not all splenic DC subpopulations in vivo. This leads to differential antigen uptake and degradation, cytokine production, cell loss, and cell death rates in various DC subpopulations. The data suggest that these effects might be caused directly by injection of Yops into DCs and indirectly by affecting the homeostasis of CD4(+) and CD8α(+) DCs. These events may contribute to reduced T-cell proliferation and immune evasion of Ye.
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Linfocitos T CD4-Positivos/inmunología , Células Dendríticas/inmunología , Células Dendríticas/microbiología , Evasión Inmune/inmunología , Factores de Virulencia/metabolismo , Yersiniosis/inmunología , Yersinia enterocolitica/inmunología , Proteínas Adaptadoras del Transporte Vesicular/fisiología , Animales , Presentación de Antígeno , Western Blotting , Linfocitos T CD4-Positivos/microbiología , Citocinas/metabolismo , Células Dendríticas/metabolismo , Femenino , Citometría de Flujo , Técnica del Anticuerpo Fluorescente , Técnicas para Inmunoenzimas , Activación de Linfocitos , Ratones , Ratones Endogámicos C57BL , Bazo/citología , Bazo/inmunología , Bazo/microbiología , Receptor Toll-Like 4/fisiología , Factores de Virulencia/genética , Yersiniosis/metabolismo , Yersiniosis/microbiologíaRESUMEN
The effect of multimodal imagery on anxiety and perceived stress was investigated in 46 male tennis players (16 to 18 yr.; M = 17.0, SD = 0.8). There were two groups, a multimodal imagery group and a control imagery group. The quasi-experimental design included pre- and post-treatment administrations of the 27-item Competitive State Anxiety Inventory to assess anxiety and the Perceived Stress Scale to assess stress. Analysis showed a statistically significant multivariate difference between the multimodal imagery and control groups on measures of cognitive anxiety, self-confidence, and perceived stress. Imagery plays a role in handling some specific types of anxiety and stress.
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Ansiedad/psicología , Atletas/psicología , Terapia Cognitivo-Conductual/métodos , Autoimagen , Estrés Psicológico/psicología , Tenis/psicología , Adolescente , Ansiedad/terapia , Humanos , Imaginación/fisiología , Masculino , Estrés Psicológico/terapia , Encuestas y CuestionariosRESUMEN
The aim of this study was to investigate the influence of ad libitum access to compound feeds rich in fibre (ante partum (a.p.) and peri partum) on the DM intake, body mass development and performance of sows as well as excretion of Clostridium (C.) perfringens via sows' faeces. From day 109 (d-7) of gestation, 25 of 48 sows (23 considered as control) received access to one of two different high-fibre pellets from d-7 until the second day post partum (p.p.) (d2) (fibre groups (FG) 1 and 2) in additional to a lactation diet. The additional DM intake of the high-fibre pellets a.p. was 2.13 ± 1.15 kg in FG 1 and 3.14 ± 0.68 kg in FG 2. This led to higher DM intake in the first lactation week and significantly lower losses of weight and back fat thickness during lactation. The bacterial counts of C. perfringens in sows' faeces directly p.p. were 10 times lower in FG 1 and 100 times lower in FG 2 compared to the controls. High amounts of fibre led to higher DM intake throughout lactation, which had beneficial effects on sows' body conditions. It seems that high fibre intake influenced the excretion of C. perfringens at parturition, which could improve the health of newborns.
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Background Multiple sclerosis (MS) is a chronic disabling disease that is associated with negative effects on health-related quality of life (HRQoL) due to reduced physical and psychosocial functioning. Cladribine tablets 10 mg (3.5 mg/kg cumulative dose over 2 years) have been approved for the treatment of adult patients with highly active relapsing multiple sclerosis (RMS). The ongoing CLARIFY-MS study (NCT03369665; EudraCT number: 2017-002632-17) aims to assess the effect of cladribine tablets 3.5 mg/kg on HRQoL of patients with highly active RMS. Objective To report on the design of the CLARIFY-MS study, baseline patient characteristics, and results of a pre-planned interim analysis focusing on treatment satisfaction, safety, and tolerability that includes all data reported till 6 months after start of treatment. Methods The CLARIFY-MS study is a 2-year, open-label, single-arm, prospective, multicenter, phase IV study. Eligible patients with highly active RMS were assigned to receive cladribine tablets 3.5 mg/kg over 2 years. Treatment satisfaction was assessed using the Treatment Satisfaction Questionnaire for Medication (TSQM, v1.4; scale range from 0 to 100, higher values indicating higher satisfaction). Safety assessments, including occurrence of treatment-emergent adverse events (TEAEs; any adverse event reported after drug administration), serious adverse events (SAEs), and lymphocyte counts, were summarized descriptively. Results A total of 482 patients from 85 sites in Europe were treated with cladribine tablets. Mean patient age was 37.4 years, 338 (70.1%) were women, median EDSS was 2.5, and 345 (71.6%) were prior users of disease-modifying therapy (DMT). During the first 6 months after the start of treatment, and before reaching the full dose of cladribine tablets, mean TSQM global satisfaction score for the overall population was 70.4 (standard deviation, ± 18.48). The side effects score was 91.9 (± 17.68), convenience scored 86.6 (± 13.57), and effectiveness was 65.8 (± 21.14). A total of 275 patients (57.1%) reported at least one TEAE and 9 patients (1.9%) had a SAE. The majority of observed lymphopenia cases were of grade 1 or 2; 33 (6.8%) of the total study cohort had grade 3 lymphopenia, and no grade 4 lymphopenia was reported. Conclusion Patients reported high treatment satisfaction (TSQM) with cladribine tablets in this pre-planned interim analysis at 6 months. Few serious, and no unexpected, adverse events were reported, and there were no instances of grade 4 lymphopenia over the first 6 months. These preliminary data indicate good tolerability and convenience of administration of cladribine tablets in patients with highly active RMS.
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Esclerosis Múltiple Recurrente-Remitente , Esclerosis Múltiple , Adulto , Cladribina/efectos adversos , Femenino , Humanos , Inmunosupresores/efectos adversos , Esclerosis Múltiple Recurrente-Remitente/tratamiento farmacológico , Recurrencia Local de Neoplasia , Satisfacción del Paciente , Satisfacción Personal , Estudios Prospectivos , Calidad de Vida , ComprimidosRESUMEN
BACKGROUND AND OBJECTIVES: The onset of action for high-efficacy immunotherapies in multiple sclerosis (MS) is an important parameter. This study (MAGNIFY-MS) evaluates the onset of action of cladribine tablets by observing changes in combined unique active (CUA) MRI lesion counts during the first 6 months of treatment in patients with highly active relapsing MS. METHODS: MRI was performed at screening, baseline, and at months 1, 2, 3, and 6 after initiating treatment with cladribine tablets 3.5 mg/kg. CUA lesion counts, defined as the sum of T1 gadolinium-enhancing (Gd+) lesions and new or enlarging active T2 lesions (without T1 Gd+), were compared between postbaseline and the baseline period and standardized to the period length and the number of MRIs performed. RESULTS: Included in this analysis were 270 patients who received ≥1 dose of cladribine tablets. After treatment initiation, significant reductions in mean CUA lesion counts were observed from month 1 onward compared with the baseline period (-1.193 between month 1 and month 6, -1.500 between month 2 and month 6, and -1.692 between month 3 and month 6; all p < 0.0001). Mean T1 Gd+ lesion counts were decreased from month 2 onward compared with baseline (-0.857 at month 2, -1.355 at month 3, and -1.449 at month 6; all p < 0.0001), whereas the proportion of patients without any CUA lesions increased from 52.0% between month 1 and month 6 to 80.5% between month 3 and month 6. DISCUSSION: Findings suggest an early onset of action for cladribine tablets, with an increasing reduction in active MRI lesions over time. TRIAL REGISTRATION INFORMATION: NCT03364036; Date registered: December 06, 2017. CLASSIFICATION OF EVIDENCE: Using frequent MRI assessments of the brain over the first 6 months of the MAGNIFY-MS study (NCT03364036), we aimed to determine the onset of action of cladribine tablets 3.5 mg/kg in adult patients with highly active relapsing MS. This study provides Class IV evidence that, in such patients, treatment with cladribine tablets is associated with an early onset of action with reductions in active MRI lesion counts from month 2 (day 60) onward, with an increasing reduction in such lesions over time.
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Cladribina , Esclerosis Múltiple , Adulto , Cladribina/farmacología , Humanos , Inmunosupresores/efectos adversos , Imagen por Resonancia Magnética , Esclerosis Múltiple/diagnóstico por imagen , Esclerosis Múltiple/tratamiento farmacológico , Comprimidos/uso terapéuticoRESUMEN
INTRODUCTION: In the Cladribine Tablets Treating Multiple Sclerosis Orally (CLARITY) study, cladribine tablets significantly reduced relapse rates and improved findings on magnetic resonance imaging versus placebo in patients with relapsing multiple sclerosis. In the CLARITY Extension study, treatment with cladribine tablets for 2 years followed by placebo for 2 years produced similar clinical benefits to 4 years of cladribine tablets. The objective of this exploratory post hoc analysis was to evaluate long-term disease stability (assessed by the Expanded Disability Status Scale [EDSS] score) after treatment with cladribine tablets. METHODS: Patients enrolled into CLARITY Extension who were previously randomized to cladribine tablets 3.5 mg/kg in the CLARITY study were included in this post hoc analysis. Two treatment groups were investigated-patients randomized to cladribine tablets 3.5 mg/kg in CLARITY and thereafter randomized to placebo in CLARITY Extension (the CP3.5 group) or to cladribine tablets 3.5 mg/kg in CLARITY Extension (the CC7 group). In each treatment group, EDSS scores at 6-month intervals, EDSS score improvement/worsening each year, and time to 3- and 6-month confirmed EDSS progression were assessed from CLARITY baseline over 5 years of follow-up (including a variable bridging interval between studies). All analyses are descriptive, and no statistical comparisons were performed for between-treatment group differences. RESULTS: The median (95% confidence interval [CI]) EDSS score for patients in the CP3.5 group at 5 years was 2.5 (2.0-3.5) compared with 3.0 (2.5-3.5) at baseline. In the CC7 group, median EDSS score (95% CI) at 5 years was 2.0 (2.0-3.0) compared with 2.5 (2.5-3.0) at baseline. During year 5 for the CP3.5 group, and based on changes in minimum score each year, EDSS score stability was observed in 53.9% of patients, improvement in 21.3%, and worsening in 24.7%. In the CC7 group, EDSS score remained stable in 66.1%, improved in 18.1%, and worsened in 15.8% of patients. Over 70% of patients in both treatment groups did not show 3- or 6-month confirmed EDSS progression at 5 years from CLARITY baseline. CONCLUSIONS: These findings confirm long-term beneficial effects on disability afforded by either the recommended dose of cladribine tablets over 4 years (cumulative dose, 3.5 mg/kg) or a higher cumulative dose. TRIAL REGISTRATION: ClinicalTrials.gov NCT00213135 (CLARITY); NCT00641537 (CLARITY Extension).
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Esclerosis Múltiple Recurrente-Remitente , Esclerosis Múltiple , Cladribina/uso terapéutico , Humanos , Inmunosupresores/uso terapéutico , Esclerosis Múltiple Recurrente-Remitente/tratamiento farmacológico , Recurrencia , ComprimidosRESUMEN
The purpose was three-fold: (1) to investigate the effect of baseline, precompetition, and postcompetition stress on salivary cortisol levels in top-ranking Brazilian wrestlers (N = 17) participating in a national competition; (2) to estimate correlations among three stress measures (perceived stress, salivary cortisol, and physiological stress reaction); and (3) to compare cortisol concentrations between losers and winners. Salivary cortisol was collected at baseline, pre-, and postcompetition. Physiological stress reaction and perceived stress scores were measured just before warm-up for the competition. Analysis showed a significant main effect for testing time. Correlations among the stress measures were not significant. Analysis of covariance between the winners (n = 10) and the losers (n = 7) was also not significant. Salivary cortisol concentrations increased after the intense exercise of competition. The wrestlers did not perceive any physiological effects.
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Nivel de Alerta/fisiología , Rendimiento Atlético/fisiología , Conducta Competitiva/fisiología , Emociones/fisiología , Hidrocortisona/sangre , Lucha/fisiología , Logro , Adolescente , Adulto , Brasil , Humanos , Masculino , Saliva/química , Estadística como Asunto , Estrés Fisiológico/fisiología , Adulto JovenRESUMEN
Liquid feed is susceptible to microbiological growth. Yeasts are said to cause sudden death in swine due to intestinal gas formation. As not all animals given high yeast content feed fall ill, growth and gas formation potential at body temperature were investigated as possible causally required properties. The best identification method for these environmental yeasts should be tested beforehand. Yeasts derived from liquid diets without (LD - S) and liquid diets with maize silage (LD + S) were examined biochemically (ID32C-test) and with MALDI-TOF with direct smear (DS) and an extraction method (EX). Growth temperature and gas-forming potential were measured. With MALDI-EX, most yeast isolates were identified: Candida krusei most often in LD - S, and C. lambica most often in LD + S, significantly more than in LD - S. Larger colonies, 58.75% of all yeast isolates, were formed at 25 °C rather than at 37 °C; 17.5% of all isolates did not grow at 37 °C at all. Most C. krusei isolates formed high gas amounts within 24 h, whereas none of the C. lambica, C. holmii and most other isolates did. The gas pressure formed by yeast isolates varied more than tenfold. Only a minority of the yeasts were able to produce gas at temperatures common in the pig gut.
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Introducing high numbers of lactic acid bacteria into the gastrointestinal tract of pigs via fermented liquid feed (FLF) could have an impact on intestinal bacterial ecosystems. Twenty piglets were allocated into four groups and fed a botanically identical liquid diet that was offered either non-fermented (twice), fully fermented or partially fermented but supplemented with 40% of non-fermented coarse cereals. Microbiota studies were performed on the small and large intestine digesta and faecal samples. A 16S rRNA gene amplification was performed within the hypervariable region V4 and sequenced with the Illumina MiSeq platform. R (version 3.5.2) was used for the statistical analyses. The digesta of the small intestines of pigs fed FLF were dominated by Lactobacillaceae (relative abundance up to 95%). In the colonic contents, the abundance of Lactobacillaceae was significantly higher only in the pigs fed the FLF supplemented with non-fermented coarse cereals. Additionally, the digesta of the small and large intestines as well as in the faeces of the pigs fed the FLF supplemented with non-fermented coarse cereals were significantly enriched for two operational taxonomic units (OTUs) belonging to the genus Lactobacillus and Bifidobacterium. The FLF supplemented with non-fermented coarse cereals had probiotic and prebiotic-like impacts on the intestinal and faecal bacterial composition of pigs.
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Anthropomorphism of dogs has affected feeding and the choice of components present in diets for dogs. Conflicting trends are present: raw or vegetarian appear more prevalent. Animal-derived proteins seem to have unfavourable impacts on intestinal microflora by decreasing the presence of Bacteroidetes. This preliminary study evaluates whether effects of diets with animal proteins on intestinal microbiota can be compensated by the addition of certain carbohydrates to dog diet. Eight female beagles were included in a cross-over study and fed a vegetarian diet or the same diet supplemented with feather meal (2.7%) and either 20% of cornmeal, fermented or non-fermented rye (moisture content of the diets about 42%). A 16S rRNA gene amplification was performed within the hypervariable region V4 on faecal samples and sequenced with the Illumina MiSeq platform. The Firmicutes/Bacteroidetes ratio tended to shift to the advantage of Firmicutes when feather meal and cornmeal were added (Firmicutes/Bacteroidetes ratio of 5.12 compared to 2.47 when offered the vegetarian diet) and tended to switch back to the advantage of Bacteroidetes if rye: fermented (2.17) or not (1.03) was added. The addition of rye might have the potential to compensate possible unfavourable effects of diets with animal proteins on intestinal microbiota of dogs.
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Carriage of resistant bacteria and spread of antimicrobial resistance (AMR) in the environment through animal manure pose a potential risk for transferring AMR from poultry and poultry products to the human population. Managing this risk is becoming one of the most important challenges in livestock farming. This study focused on monitoring the prevalence of multi-drug resistance (MDR) bacteria and development of AMR depending on flooring. In two experiments (2 × 15,000 birds), broilers were always divided in two different stables. In the control group, the entire floor pen was covered with litter material and in the experimental group, the flooring system was partly modified by installing elevated slat platforms equipped with water lines and feed pans. Over the whole fattening period, excreta and manure samples were taken (days 2, 22, and 32). In total, 828 commensal E. coli isolates were collected. The development and prevalence of resistance against four different antibiotic classes (quinolones, ß-lactams, tetracyclines, and sulfonamides) were examined by using broth microdilution. At the end of the trials, the amount of manure per square metre was twice as high below the elevated platforms compared to the control group. Approximately 58% of E. coli isolates from excreta showed resistance against at least one antibacterial agent at day 2. During and at the end of the fattening period, resistant E. coli isolates at least against one of the four antibacterial agents were observed in excreta (46 and 46%, respectively), and manure samples (14 and 42%, respectively), despite the absence of antibacterial agent usage. In spite of less contact to manure in the experimental group, the prevalence of resistant E. coli isolates was significantly higher. Birds preferred the elevated areas which inevitably led to a local high population density. Animal-to-animal contact seems to be more important for spreading antimicrobial resistant bacteria than contact to the litter-excreta mixture. Therefore, attractive areas in poultry housing inducing crowding of animals might foster transmission of AMR. In poultry farming, enrichment is one of the most important aims for future systems. Consequently, there is a need for keeping birds not carrying resistant bacteria at the start of life.
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The transition from breast milk to solid feed is a dramatic change in the nutrition of piglets, frequently necessitating antibiotic treatment. In efforts to reduce the use of antibiotics, dietetic concepts based on natural feed additives are becoming more and more important. In the present study, experiments were carried out with 15 rearing piglets (days 28-56) divided into three groups that were offered different diets (Ctr [0% peat]; H1.5 [1.5% peat]; and H3.0 [3.0% peat] based on a commercial weaner recipe; all ~178 g CP, 13.7 MJ ME, 13.3 g Lys, as-fed). The contents of cecal and colon digesta were removed at necropsy. The gas formation (4 h) in colon digesta was measured using in vitro batch fermenters. For microbiome studies, 16S rRNA amplification was performed within the hypervariable region V 4 and sequenced with Illumina MiSeq platform. DNA read mapping and statistical analysis were performed using QIIME (version 1.8.0), MicrobiomeAnalyst, RStudio, and SAS Enterprise Guide. The mean body weight of the animals at the end of the trial did not show statistical differences (in kg: Ctr: 26.1 ± 4.85, H1.5: 28.5 ± 3.41, H3.0: 26.2 ± 4.92). The daily weight gains were high for this age (in g/day; Ctr: 607 ± 157; H1.5: 692 ± 101; H3.0: 615 ± 113) and the feed to gain ratio low (in kg/kg; Ctr: 1.538; H1.5: 1.462; H3.0: 1.462). Concentrations of short-chain fatty acids in the cecal content were significantly lower when peat was used (mmol/kg wet weight; Ctr: 173 ± 30.0; H1.5:134 ± 15.0; H3.0:133 ± 17.3). Numerical differences were found in the gas formation (in mL gas per 10 mL batch in 4 h; Ctr: 7.9 ± 2.2; H1.5: 7.4 ± 2.4; H3.0: 6.6 ± 1.1). The microbiome analyses in the cecal content showed significantly higher values for alpha diversity Chao 1 index for samples from the control group. Significant differences were found for bacterial relative abundance for Tenericutes at phylum level and Mollicutes at class level (p < 0.05) in cecal microbiota. Therefore, there was initial evidence that peat influences intestinal microflora causing a shift in the overall concentration of fermentation products in both, the cecal and the colon content.
RESUMEN
Campylobacter jejuni is the most common bacterial cause of foodborne zoonosis in the European Union. Infections are often linked to the consumption and handling of poultry meat. The aim of the present study was to investigate the caecal microbiota of birds infected with C. jejuni at different ages. Therefore, a total of 180 birds of the laying hybrid Lohmann Brown-Classic were housed in 12 subgroups of 15 animals each in three performed repetitions. Three birds per subgroup were experimentally infected with C. jejuni at an age of about 21 days and about 78 days (4.46 ± 0.35 log10 CFU/bird). Twenty-one days after experimental infection, microbiome studies were performed on 72 caecal samples of dissected birds (three primary infected and three further birds/subgroup). Amplification within the hypervariable region V 4 of the 16S rRNA gene was performed and sequenced with the Illumina MiSeq platform. Statistical analyses were performed using SAS® Enterprise Guide® (version 7.1) and R (version 3.5.2). Both factors, the experimental replication (p < 0.001) and the chickens' age at infection (p < 0.001) contributed significantly to the differences in microbial composition of the caecal samples. The factor experimental replication explained 24% of the sample's variability, whereas the factor age at infection explained 14% thereof. Twelve of 32 families showed a significantly different count profile between the two age groups, whereby strongest differences were seen for seven families, among them the family Campylobacteraceae (adjusted p = 0.003). The strongest difference between age groups was seen for a bacterial species that is assigned to the genus Turicibacter which in turn belongs to the family Erysipelotrichaceae (adjusted p < 0.0001). Correlation analyses revealed a common relationship in both chicken ages at infection between the absolute abundance of Campylobacteraceae and Alcaligenaceae, which consists of the genus Parasutterella. In general, concentrations of particular volatile fatty acids (VFA) demonstrated a negative correlation to absolute abundance of Campylobacteraceae, whereby the strongest link was seen for n-butyrate (-0.51141; p < 0.0001). Despite performing consecutive repetitions, the factor experimental replication contributed more to the differences of microbial composition in comparison to the factor age at infection.
RESUMEN
The objective of this study was to evaluate the effects of different types of flooring designs on antimicrobial resistance in commensal Escherichia coli from turkeys treated with enrofloxacin. Two trials were performed with same feed, housing conditions and fattening duration, but with different flooring designs. Furthermore, the first trial was performed in an unchanged environment and the second trial in a changed environment. The flooring designs of the pens were assigned to four groups; G1 - entire floor pen covered with litter, G2 - floor pen with heating, G3 - partially slatted flooring including an area that was littered, G4 - fully slatted flooring with a sand bath. Enrofloxacin was given at days 10-14 via drinking water. The changed environment in the second trial was achieved by moving the animals to new pens with the respective same conditions as previously after antimicrobial administration at day 15. A total of 576 E. coli were isolated from cloacal swab and poultry manure samples. Sample collection was done before the treatment, after the treatment and at the end of the trials at day 35. The resistance of isolates to enrofloxacin and ampicillin was determined using broth microdilution A single treatment with enrofloxacin reduced the proportion of samples with susceptible E. coli isolates significantly in all flooring designs. Overall, frequencies of enrofloxacin resistance were significantly different between the unchanged and changed environment. At the same time, the proportion of ampicillin-resistant isolates increased in the first trial, although no ampicillin was applied in this study.
Asunto(s)
Farmacorresistencia Bacteriana , Infecciones por Escherichia coli/veterinaria , Escherichia coli/efectos de los fármacos , Pisos y Cubiertas de Piso , Vivienda para Animales , Simbiosis , Pavos/microbiología , Animales , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Enrofloxacina , Escherichia coli/aislamiento & purificación , Infecciones por Escherichia coli/tratamiento farmacológico , Infecciones por Escherichia coli/microbiología , Fluoroquinolonas/administración & dosificación , Fluoroquinolonas/uso terapéutico , Pruebas de Sensibilidad Microbiana , Enfermedades de las Aves de Corral/tratamiento farmacológicoRESUMEN
Gaining knowledge about the spread of resistance against antibacterial agents is a primary challenge in livestock farming. The purpose of this study was to test the effect of double antibiotic treatment (at days 10â»14 and days 26â»30) with enrofloxacin or solely environmental exposition (identical times, directly into the litter) on resistance against antibacterial agents in commensal Escherichia coli in comparison with the control (without treatment), depending on different flooring. A total of 720 Big 6 turkeys participated in three trials. Four different flooring designs were examined: An entire floor pen covered with litter, a floor pen with heating, a partially slatted flooring including 50% littered area, and a fully slatted flooring with a sand bath. A total of 864 Escherichia coli isolates were obtained from cloacal swabs and poultry manure samples at days 2, 9, 15, 21, and 35. The broth microdilution method (MIC) was used to determine the resistance of isolates to enrofloxacin and ampicillin. A double antibiotic treatment with enrofloxacin reduced the proportion of susceptible Escherichia coli isolates significantly in all flooring designs. Simulation of water losses had no significant effect, nor did the flooring design. Ampicillin-resistant isolates were observed, despite not using ampicillin.