Black Garlic and Thiosulfinate-Enriched Extracts as Adjuvants to Ceftriaxone Treatment in a Rat Peritonitis Model of Sepsis.

To date, there have been no new drugs or adjuvants able to decrease both morbidity and mortality in the context of sepsis and septic shock. Our objective was to evaluate the use of thiosulfinate-enriched Allium sativum and black garlic extracts as adjuvants in the management of sepsis. An experimental in vivo study was carried out with male Sprague-Dawley® rats. Animals were randomized in four treatment groups: antibiotic (ceftriaxone) treatment (group I), ceftriaxone plus thiosulfinate-enriched extract (TASE, group II), ceftriaxone plus thiosulfinate-enriched extract and black garlic extracts (TASE + BGE, group III), and ceftriaxone plus black garlic extract (BGE, group IV). All animals were housed and inoculated with 1 × 1010 CFU/15 mL of intraperitoneal Escherichia coli ATCC 25922. Subsequently, they received a daily treatment according to each group for 7 days. Clinical, analytical, microbiological, and histopathological parameters were evaluated. Statistically significant clinical improvement was observed in rats receiving garlic extracts in weight (groups II and III), ocular secretions, and piloerection (group IV). Moreover, less liver edema, vacuolization, and inflammation were observed in groups receiving adjuvant support (groups II, III, and IV). When comparing interleukins 24 h after bacteria inoculum, we found statistically significant differences in TNF-alpha levels in groups receiving BGE (groups III and IV, p ≤ 0.05). Blood and peritoneal liquid cultures were also analyzed, and we detected a certain level of Enterococcus faecalis in peritoneal cultures from all treatment groups and less bacteria presence in blood cultures in rats receiving garlic extracts (groups II, III, and IV). In conclusion, TASE and BGE could be promising nutraceutical or medicinal agents as coadjuvants in the treatment of sepsis because of its effects in modulating the inflammatory response.

Prevalence and risks factors associated with ESBL-producing faecal carriage in a single long-term-care facility in Spain: emergence of CTX-M-24- and CTX-M-27-producing Escherichia coli ST131-H30R.

OBJECTIVES: To address the faecal carriage prevalence of antibiotic-multiresistant bacteria and associated risk factors in a public long-term care facility (LTCF). METHODS: A prospective study in a single government-funded LTCF of 300 residents in Ciudad Real, Spain. Residents' clinical and demographic data were collected, as well as recent antibiotic consumption in the institution. Each participant contributed a rectal swab, which was plated on selective and differential-selective media. Colonies were identified by MALDI-TOF and ESBL production was confirmed by the double-disc synergy method, with characterization of the molecular mechanism by PCR. Isolates were typed by PFGE and submitted for ST131 screening by PCR. RESULTS: Faecal carriage of ESBL-producing Enterobacterales was detected in 58 (31%) of 187 participants and previous infection by MDR bacteria was identified as a risk factor. The genes characterized were: blaCTX-M-15 (40.6%); blaCTX-M-14 (28.8%); blaCTX-M-27 (13.5%); and blaCTX-M-24 (10.1%). Some 56.4% of the isolates were grouped into the E. coli ST131 clone; 70.9% of these corresponded to the O25b serotype, 51.6% of them to Clade C1 (H30) and 12.9% to Clade C2 (H30Rx). Clade C1 isolates were mostly C1-M27, whereas the C2 sublineage was mainly related to the production of CTX-M-15. ST131-CTX-M-24 isolates (n = 6) corresponded to Clade A with serotype O16. CONCLUSIONS: A high prevalence of ESBL-producing Enterobacterales faecal carriage has been detected in a single LTCF, highlighting the emergence of ST131 Clade A-M24 and Clade C1-M27 lineages.

Desarrollo de modelo experimental animal de peritonitis bacteriana / Development of animal experimental model for bacterial peritonitis

OBJETIVO: Desarrollar un modelo de sepsis abdominal en animal de experimentación. MATERIAL Y MÉTODOS: Se utilizan ratas Sprague-Dawley(R), machos de 5 semanas con pesos entre 270-280 g en el momento de la inoculación (N=39). Inicialmente se realiza un estudio piloto (N=9), distribuyéndolas en 3 grupos (3/3/3) con inóculo de 1cc de Escherichia coli ATCC 25922 intraperitoneal en concentraciones de 10E8, 10E9 y 10E10 UFC. En un segundo estudio (N=6) con distribución en dos grupos (3/3) se utilizan 1cc una concentración de E. coli 10E10 UFC que se diluyen en 10 y 15 ml de agua destilada para su inoculación. Por último se inicia un ensayo experimental con aleatorización de 24 ratas en tres grupos de tratamiento tras la infección intraperitoneal: Grupo I con suero fisiológico (N=6), Grupo II con antibiótico (ceftriaxona) (N=9), Grupo III con antibiótico más adyuvante (ceftriaxona más alicina) (N=9). Se realizan muestras microbiológicas de sangre y líquido peritoneal, así como estudio histopatológico de órganos intraperitoneales (hígado, diafragma y peritoneo). RESULTADOS: Se observa muerte en el 100% de las ratas infectadas con la concentración de E. coli 10E10 UFC con la dilución de 15 ml de agua destilada y sin antibiótico. El hemocultivo y cultivo de líquido peritoneal es positivo a la misma cepa en todas ellas. Se observa la formación de abscesos en la superficie del hígado e infiltración por polimorfonucleares en los tejidos. CONCLUSIÓN: Se establece que la dosis letal de E. coli es 10E10 UFC diluida en 15 ml agua destilada en inyección intraperitoneal

OBJECTIVE: The aim of the study was to develop a model of abdominal sepsis in the experimental animal. MATERIAL AND METHODS: Sprague-Dawley male rats of 5 weeks (N=39) were used. Initially, a pilot study (N = 9) was performed and distributed in 3 groups with 1cc inoculum of Escherichia coli ATCC 25922 intraperitoneally at concentrations of 10E8, 10E9 and 10E10 CFU. Subsequently, concentrations of 10E10 CFU are used in two groups of 3 rats with dilutions of 10 cc and 15 cc of distilled water respectively. Finally, a randomized trial of 24 rats was started in three treatment groups after intraperitoneal infection: Group I with physiological serum (N = 6), Group II with ceftriaxone (N = 9), Group III with ceftriaxone plus allicin (N = 9). Microbiological samples of blood and peritoneal fluid were made, as well as histopathological study of intraperitoneal organs (liver, diaphragm and peritoneum). RESULTS: Death of 100% of the rats infected with 10E10 E. coli UFC concentration with the dilution of 15 ml of distilled water and without antibiotic was oberved. The blood culture and peritoneal fluid culture was positive for the same strain in all of them. The formation of abscesses on the liver surface and polymorphonuclear infiltration in tissues were observed. CONCLUSION: The lethal dose of E. coli is 10E10 CFU diluted in 15 cc distilled water by intraperitoneal injection

Interferon-γ production by CMV-specific CD8+ T lymphocytes provides protection against cytomegalovirus reactivation in critically ill patients.

PURPOSE: To evaluate the usefulness of the secretion of interferon-γ (IFNγ) by cytomegalovirus (CMV)-specific CD8+ T cells to determine the risk of CMV reactivation in critically ill non-immunosuppressed patients. METHODS: Two-center prospective cohort study including critically ill non-immunosuppressed CMV-seropositive patients admitted between December 2012 and March 2013. The incidence of CMV reactivation by polymerase chain reaction (real-time PCR) in plasma was investigated. IFNγ secretion by CMV-specific CD8+ T lymphocytes was determined at the time of admission to the intensive care unit (ICU) by means of the QuantiFERON(®)-CMV (QF-CMV) test. Cox regression analyses were performed to investigate CMV reactivation risk factors. RESULTS: Fifty-three patients were included, of whom 13 (24.5%) presented CMV reactivation. Twenty-six patients (49.1%) were QF-CMV "reactive" (QF-CMV(R)). Of the 26 QF-CMV(R) patients, 11.5% (3/26) had CMV reactivation, whereas 37% (10/27) of QF-CMV "non reactive" patients (QF-CMV(NR)) presented reactivation (p = 0.03). By Cox regression, the presence of QF-CMV(R) at ICU admission (HR 0.09, 95% CI 0.02-0.44; p = 0.003) was associated with a decreased risk of CMV reactivation. The sensitivity, specificity, positive predictive value, and negative predictive value of QF-CMV were 77, 57, 37, and 88%, respectively. Eleven of the 53 patients (20.7%) died during the follow-up period. Mortality was more frequent in patients with CMV reactivation (6/13, 46.1 vs. 5/40, 12.5%; p = 0.015). CONCLUSIONS: In critically ill non-immunosuppressed patients, the presence of functional CMV-specific CD8+ T lymphocyte response at intensive care unit admission provides protection against CMV reactivation.

High vancomycin minimum inhibitory concentration is associated with poor outcome in patients with methicillin-susceptible Staphylococcus aureus bacteremia regardless of treatment.

We retrospectively investigated the impact of high vancomycin minimum inhibitory concentration (MIC > 2 µg/ml) on the outcome of 53 patients with bacteremia caused by methicillin-susceptible Staphylococcus aureus (MSSA). Vancomycin MIC was determined by broth microdilution according to CLSI methods. The primary outcome was 30-day all-cause mortality from the date of the first positive blood culture. The mortality rate was 22.6% (12 of 53 patients). High vancomycin MIC (odds ratio (OR) = 9.3; 95% confidence interval (95% CI) = 1.31-63.20; p = 0.027), Charlson comorbidity index ≥ 3 (OR = 10.3; 95% CI = 1.3-102.04; p = 0.03), advanced age (OR = 35.8; 95% CI = 2.3-659.2; p = 0.01), and severe sepsis (OR = 8.5; 95% CI = 1.2-61.4; p = 0.03) were associated with mortality.

[Changes in the antimicrobial susceptibility of Escherichia coli isolates from community diagnosed urinary tract infections during the period 2003-2007. Multicentre study in Castilla la Mancha (Spain)]. / Evolución del patrón de sensibilidad de Escherichia coli en infecciones del tracto urinario diagnosticadas en la comunidad durante el periodo 2003-2007. Estudio multicéntrico en Castilla la Mancha.

OBJECTIVE: To know the evolution of susceptibility patterns of Escherichia coli in patients with community diagnosed urinary tract infections (UTIs) during last years in Castilla la Mancha (Spain). METHODS: Descriptive and retrospective study performed between january 2003 and december 2007. We studied data about frequency and susceptibility of 33.651 E. coli isolates from urine cultures that were remited from primary care centres depending of 6 hospitals in Castilla la Mancha (Spain). RESULTS: Susceptibility rates of E. coli for most antibiotics decreased significantly during the 5-year period, especially for amoxicillin-clavulanic acid, cefuroxime and quinolones. Average rates of susceptibility for amoxicillin-clavulanic acid, ciprofloxacin, cefuroxime, fosfomycin and nitrofurantoin were: 86.7, 75.4, 87.3, 97.6 and 96.2%, respectively. We observed a significantly increase of E. coli isolates producing extended-spectrum betalactamases ESBLs), from 1.9% in 2003 to 4,9% in 2007 (χ² TL = 143.6, p<0.001). CONCLUSIONS: We observed a significantly reduction of E. coli susceptibility for most antibiotics and an increase of E. coli isolates producing ESBLs. Fosfomycin and nitrofurantoin are the best choices for empiric treatment. Prospective studies should be performed in the future to confirm the results of our study.

Evolución del patrón de sensibilidad de Escherichia coli en infecciones del tracto urinario diagnosticadas en la comunidad durante el periodo 2003-2007. Estudio multicéntrico en Castilla la Mancha / Changes in the antimicrobial susceptibility of Escherichia coli isolates from communitydiagnosed urinary tract infections during the period 2003-2007. Multicentre study in Castilla la Mancha (Spain)

Objetivo: Conocer la evolución del patrón de sensibilidadantimicrobiana de Escherichia coli en infecciones del tractourinario (ITUs) diagnosticadas en la comunidad durante los últimosaños en Castilla la Mancha.Métodos: Estudio descriptivo de carácter retrospectivo queabarcó desde enero de 2003 hasta diciembre de 2007. Se analizarondatos de sensibilidad de 33.651 aislados de E. coli procedentesde urocultivos remitidos desde los Centros de Atención Primariadependientes de 6 hospitales de Castilla la Mancha.Resultados: Se obtuvo una tendencia lineal significativaen la disminución de la sensibilidad de E. coli para la mayorparte de antibióticos, siendo más acusado para amoxicilinaácidoclavulánico, cefuroxima y quinolonas. Los porcentajesmedios de sensibilidad a amoxicilina-ácido clavulánico, ciprofloxacino,cefuroxima, fosfomicina y nitrofurantoína fueron:86,7, 75,4, 87,3, 97,6 y 96,2%, respectivamente. Se observó unincremento significativo de la frecuencia de cepas de E. coliportadoras de betalactamasas de espectro extendido (BLEEs),oscilando desde el 1,9% en el año 2003 hasta el 4,9% en el año2007 (χ2 TL = 143,6, p<0,001).Conclusiones: En Castilla la Mancha se está produciendoun descenso significativo de la sensibilidad de E. coli a la mayorparte de antibióticos y un incremento progresivo de las cepasportadoras de BLEEs. Fosfomicina y nitrofurantoína constituyenlas mejores opciones terapéuticas para el tratamientoempírico. Sería necesario llevar a cabo futuros trabajos de carácterprospectivo con el fin de confirmar los resultados obtenidosen el presente estudio(AU)

Objective: To know the evolution of susceptibilitypatterns of Escherichia coli in patients with communitydiagnosedurinary tract infections (UTIs) during last yearsin Castilla la Mancha (Spain).Methods: Descriptive and retrospective study performedbetween january 2003 and december 2007. We studieddata about frequency and susceptibility of 33.651E. coli isolates from urine cultures that were remitedfrom primary care centres depending of 6 hospitals inCastilla la Mancha (Spain).Results: Susceptibility rates of E. coli for most antibioticsdecreased significantly during the 5-year period,especially for amoxicillin-clavulanic acid, cefuroximeand quinolones. Average rates of susceptibility for amoxicillin-clavulanic acid, ciprofloxacin, cefuroxime, fosfomycinand nitrofurantoin were: 86,7, 75,4, 87,3, 97,6and 96,2%, respectively. We observed a significantly increaseof E. coli isolates producing extended-spectrumbetalactamases (ESBLs), from 1,9% in 2003 to 4,9% in2007 (χ2 TL = 143,6, p<0,001).Conclusions: We observed a significantly reductionof E. coli susceptibility for most antibiotics and an increaseof E. coli isolates producing ESBLs. Fosfomycinand nitrofurantoin are the best choices for empiric treatment.Prospective studies should be performed in the futureto confirm the results of our study(AU)