Optimal lameness induction model development using amphotericin B in meat goats.

Lameness continues to be a critical health and welfare concern associated with goat production. Amphotericin B (amp B) is an antimicrobial successful in inducing transient lameness for research purposes previously in livestock animals. The objectives of this study were to (1) identify which of three varying doses of amp B would be most effective in inducing lameness in meat type goats and (2) develop a facial grimace scale for goats. Lameness was produced by an intra-articular injection of amphotericin B into the left hind lateral claw distal interphalangeal joint with either a 5 mg/0.25 mL (high-low, 5 mg of amphotericin B in a volume of 0.25 mL), 5 mg/0.5 mL (high-high, 5 mg of amphotericin B in a volume of 0.5 mL), or a 2.5 mg/0.25 mL (low-low, 2.5 mg of amphotericin B in a volume of 0.25 mL). A saline treatment of 0.5 mL was used as control (0.9% sterile saline solution). Lameness response was analyzed by infrared thermography (IRT) at the induced joint, mechanical-nociception threshold (MNT), visual lameness scoring (VLS), a visual analogue scale (VAS), kinetic gait analysis (KGA), plasma cortisol (CORT), substance P (Sub P), and behavior scoring. The IRT and MNT values differed by timepoint (P ≤ 0.0001). Results from VLS showed the HL treatment was the most effective at inducing lameness (6/6 goats became lame compared to HH 4/6 and LL 2/6). At 24, 48, and 72 h, VAS scores were significantly higher when comparing HL to all other treatment groups (P = 0.0003). Both behavior observers (1 and 2) reported a significant time effect (P = 0.05), with goats exhibiting more facial grimacing at 24 h post-lameness induction. From these data, an optimal dose for a repeatable lameness induction model in goats was aquired. An effective Goat Grimace Scale (GGS) was also developed to evaluate pain responses in goats.

The Application of 3D Landmark-Based Geometric Morphometrics towards Refinement of the Piglet Grimace Scale.

Proper methods of assessment to objectively identify pain are essential for welfare improvements of piglets undergoing surgical castration on commercial farms. The Piglet Grimace Scale (PGS) is used to identify changes in facial expressions caused by acute pain in piglets undergoing tail docking and castration. However, subjective scoring methods are a concern for the validation of the PGS. The objectives of this study were to evaluate and refine the PGS through 3D landmark geometric morphometrics. Male piglets (n = 88) were randomly allocated to one of two treatments: castration and sham-castration. Piglet facial images were taken at four time points (before treatment, immediately post-treatment, 1 h post-treatment, and 4 h post-treatment) using a photogrammetry rig. Images were scored by four raters using five facial action units (FAUs): orbital tightening, ear position, temporal tension, lip contraction, and nose bulge/cheek tension. Three-dimensional facial models were generated and landmarked denoting 3 FAUs (orbital tightening, lip contraction, and nose bulge/cheek tension). Results suggest that orbital tightening and ear position may be reliable FAUs for the PGS. However, neither the PGS nor 3D landmark-based geometric morphometrics were able (both p > 0.10) to reliably identify facial indicators of pain in piglets undergoing castration.

Successful control of Serratia marcescens outbreak in a neonatal unit of a tertiary-care hospital in Spain.

OBJECTIVE: Serratia marcescens is a Gram-negative bacterium that is found in hospital environments and commonly associated with outbreaks in neonatal units. One S. marcescens isolate was detected from a bloodstream culture from a neonate in our hospital that was followed by an outbreak. The aim of this study was to describe the molecular epidemiology of a S. marcescens outbreak in the neonatal unit. METHODS: In order to investigate the outbreak, weekly surveillance rectal swabs were submitted for culture from all patients admitted in this unit from August to September 2018. Environmental samples were obtained from potential sources in September 2018. Typing of isolates was performed by pulsed field gel electrophoresis (PFGE). In addition, we studied the in vitro activity of chlorhexidine against S. marcescens. RESULTS: During this period, 146 infants were hospitalised in our neonatal unit, of which 16 patients had a S. marcescens-positive sample. A total of 36 environmental surveillance samples were collected, and one sample from a stethoscope from an incubator of a colonized baby was positive for S. marcescens. All the 18 isolates, including the isolate from the stethoscope, belonged to a single PFGE cluster. We found that very low concentrations of chlorhexidine, even with application times close to 0 achieved significant reductions in the amount of S. marcescens. CONCLUSION: A unique clone of S. marcescens caused this outbreak, including isolates from patients and from one stethoscope. The outbreak was controlled with the early implementation of specific control measures.

Pacemaker-induced endocarditis by Gordonia bronchialis.

PURPOSE: Gordonia species are known to be opportunistic human pathogens causing secondary infections. We present the second case in the world of endocarditis caused by Gordonia bronchialis and a review of all the cases of endocarditis caused by Gordonia spp. METHODS: The identification was performed by matrix-assisted desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and 16S rRNA gene sequencing were performed to confirm the identification. Antimicrobial susceptibility was performed by MIC test Strip on Mueller-Hinton agar supplemented with 5% defibrinated sheep blood according to Clinical and Laboratory Standards Institute. RESULTS: Pacemaker-induced endocarditis due to Gordonia bronchialis infection was determined in an 88-year old woman. The patient was treated with ceftriaxone and ciprofloxacin until completing 6 weeks from the pacemaker explant with a good evolution. CONCLUSION: The case presented supports the pathogenic role of Gordonia bronchialis as an opportunistic pathogen and highlights the high risk of suffering infections caused by environmental bacteria.

Successful control of Serratia marcescens outbreak in a neonatal unit of a tertiary-care hospital in Spain / Control exitoso de un brote de Serratia marcescens en la unidad neonatal de un hospital terciario en España

ObjectiveSerratia marcescens is a Gram-negative bacterium that is found in hospital environments and commonly associated with outbreaks in neonatal units. One S. marcescens isolate was detected from a bloodstream culture from a neonate in our hospital that was followed by an outbreak. The aim of this study was to describe the molecular epidemiology of a S. marcescens outbreak in the neonatal unit.MethodsIn order to investigate the outbreak, weekly surveillance rectal swabs were submitted for culture from all patients admitted in this unit from August to September 2018. Environmental samples were obtained from potential sources in September 2018. Typing of isolates was performed by pulsed field gel electrophoresis (PFGE). In addition, we studied the in vitro activity of chlorhexidine against S. marcescens.ResultsDuring this period, 146 infants were hospitalised in our neonatal unit, of which 16 patients had a S. marcescens-positive sample. A total of 36 environmental surveillance samples were collected, and one sample from a stethoscope from an incubator of a colonized baby was positive for S. marcescens. All the 18 isolates, including the isolate from the stethoscope, belonged to a single PFGE cluster. We found that very low concentrations of chlorhexidine, even with application times close to 0 achieved significant reductions in the amount of S. marcescens.ConclusionA unique clone of S. marcescens caused this outbreak, including isolates from patients and from one stethoscope. The outbreak was controlled with the early implementation of specific control measures.

ObjetivoSerratia marcescens(S. marcescens) es una bacteria gramnegativa que se encuentra en ambientes hospitalarios y comúnmente aparece asociada a brotes en unidades neonatales. En agosto de 2018 se detectó un aislado de esta bacteria a partir de un hemocultivo de un paciente neonatal en nuestro hospital. El objetivo de este estudio fue describir la epidemiología molecular del brote de S. marcescens en la unidad neonatal.MétodosCon el fin de investigar el brote, se enviaron semanalmente para cultivo muestras rectales de todos los pacientes ingresados en estas unidades de agosto a septiembre de 2018. Asimismo, se obtuvieron muestras ambientales de potenciales orígenes del brote en septiembre de 2018. Los aislados se genotipificaron mediante electroforesis de campo pulsado (PFGE).ResultadosDurante este período, 146 lactantes fueron hospitalizados en nuestra unidad neonatal, de los cuales 16 pacientes tenían una muestra positiva para S. marcescens. Se recogieron un total de 36 muestras de vigilancia ambiental. Una muestra de un estetoscopio de una incubadora de un bebé colonizado resultó positiva para S. marcescens. Los 18 aislamientos, incluido el aislado de la muestra ambiental, pertenecían a un solo patrón de PFGE. Se realizaron experimentos in vitro con clorhexidina y se encontró que concentraciones muy bajas incluso con tiempos de aplicación cercanos a 0 lograban reducciones significativas en la cantidad de S. marcescens.ConclusiónEstos datos confirmaron un único clon de S. marcescens en la unidad neonatal con aislamientos tanto de pacientes como ambientales. La implementación temprana de medidas de control específicas fue eficaz para limitar la transmisión nosocomial.

Pacemaker-induced endocarditis by Gordonia bronchialis / Endocarditis del marcapasos debido a infección por Gordonia bronchialis

PurposeGordonia species are known to be opportunistic human pathogens causing secondary infections. We present the second case in the world of endocarditis caused by Gordonia bronchialis and a review of all the cases of endocarditis caused by Gordonia spp.MethodsThe identification was performed by matrix-assisted desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and 16S rRNA gene sequencing were performed to confirm the identification. Antimicrobial susceptibility was performed by MIC test Strip on Mueller-Hinton agar supplemented with 5% defibrinated sheep blood according to Clinical and Laboratory Standards Institute.ResultsPacemaker-induced endocarditis due to Gordonia bronchialis infection was determined in an 88-year old woman. The patient was treated with ceftriaxone and ciprofloxacin until completing 6 weeks from the pacemaker explant with a good evolution.ConclusionThe case presented supports the pathogenic role of Gordonia bronchialis as an opportunistic pathogen and highlights the high risk of suffering infections caused by environmental bacteria.

ObjetivoLas especies de Gordonia son patógenos humanos oportunistas que causan infecciones secundarias. Presentamos el segundo caso en el mundo de endocarditis causada por Gordonia bronchialis, así como una revisión de todos los casos de endocarditis causados por Gordonia spp.MétodosLa identificación fue realizada mediante espectrometría de masas MALDI-TOF MS, y se confirmó mediante secuenciación del gen 16S rRNA. La susceptibilidad antimicrobiana se realizó mediante tiras reactivas MIC en agar Müller-Hinton suplementado con un 5% de sangre ovina desfibrinada, conforme al Clinical and Laboratory Standards Institute (CLSI).ResultadosLa endocarditis del marcapasos debido a infección por Gordonia bronchialis se encontró en una mujer de 88 años. La paciente fue tratada con ceftriaxona y ciprofloxacina hasta completar el periodo de 6 semanas desde el explante del marcapasos, con buena evolución.ConclusiónEste caso respalda el rol patogénico de Gordonia bronchialis como patógeno oportunista, subrayando el alto riesgo de padecer infecciones causadas por bacterias ambientales.

Surgical Castration Changes Struggle Behavior and Vocalizations in Male Piglets.

This study assessed acute pain in piglets during castration through behavioral indicators. Piglets (n=88) were randomly allocated to one of two treatments: surgical castration and sham-castration. Within 24 hours after birth, identical castration procedures were followed for both treatment groups, except sham piglets were not castrated. Struggle behavior (curl ups, leg kicks, and body flailing) and vocalization (duration and peak frequency) were analyzed during the castration procedure. Castrated piglets kicked more frequently than sham piglets (28.8 vs. 21.3 kicks/min, SE = 0.09; P = 0.02). Additionally, 51.2% of castrated piglets displayed body flailing, whereas only 4.4% of sham piglets displayed the same behavior (P = 0.03). Castrated piglets responded with more high frequency (≥1,000 Hz) calls than sham piglets (23.6 vs. 18.6 calls/min, SE = 0.26; P = 0.04) and high frequency calls tended to be of longer duration for castrated piglets (0.45 vs. 0.27 sec/call, SE = 0.04; P = 0.08). Results indicate that surgical castration increased the frequency of leg kicks, body flailing, and high frequency calls compared to sham-castration, suggesting these may be useful behavioral indicators of acute pain in piglets.

Effect of bupivacaine liposome suspension administered as a cornual nerve block on indicators of pain and distress during and after cautery dehorning in dairy calves.

Dehorning is performed on a high percentage of dairies worldwide. Concern about the negative effect of dehorning on animal welfare has contributed to the development of new guidelines that require the use of pain management at the time of disbudding in the United States. However, livestock producers are limited in how to address this requirement due to a lack of (1) approved analgesic drugs, (2) analgesic options that control pain for an extended duration, and (3) analgesic formulations that are practical for producers to administer. The objective of this study was to evaluate the effectiveness of bupivacaine liposome suspension, a novel, long-acting, local anesthetic formulation administered as a nerve block at dehorning, compared with current industry standard analgesic approaches using lidocaine nerve blocks alone or in combination with the nonsteroidal anti-inflammatory drug meloxicam. Fifty male Holstein calves, 10 to 14 wk of age, were enrolled and randomly assigned to 1 of 5 treatment groups before cautery dehorning as follows: (1) bupivacaine liposome suspension block, oral placebo (BUP); (2) lidocaine block, oral placebo (LID); (3) lidocaine block, oral meloxicam (1 mg/kg of body weight; LID + MEL); (4) saline block, oral placebo (CON); and (5) saline block, oral placebo, sham dehorn (SHAM). Biomarkers were collected from 0 to 120 h postdehorning and included infrared thermography, mechanical nociceptive threshold (MNT), pressure mat gait analysis, chute defense and behavior scoring, and blood sampling for serum cortisol and prostaglandin E2 metabolites. Responses were analyzed using repeated measures with calf nested in treatment designated as a random effect, and treatment, time, and their interaction designated as fixed effects. At 2 h postdehorning, the BUP group had a higher MNT compared with the CON group. Furthermore, at 24 h postdehorning, the BUP group had a higher MNT compared with the LID group. Gait distance differed significantly between treatment groups; the CON, LID, and LID + MEL groups had an increased gait distance relative to the SHAM group. The CON group exhibited a higher chute defense behavior score during the dehorning procedure compared with all other treatments. Furthermore, the CON group exhibited more ear flicks than the BUP and LID + MEL groups postdehorning. At 4 h and 24 h after dehorning, the LID + MEL group had a lower average prostaglandin E2 metabolites concentration compared with all other treatment groups. These data showed that administration of bupivacaine liposome suspension as a cornual nerve block at the time of dehorning was as effective at controlling pain as a multimodal approach of lidocaine and meloxicam.

Effect of bupivacaine liposome suspension administered as a local anesthetic block on indicators of pain and distress during and after surgical castration in dairy calves.

Castration is a routine procedure performed on beef and dairy operations in the United States. All methods of castration cause behavioral, physiologic, and neuroendocrine changes associated with pain. The American Veterinary Medical Association and the American Association of Bovine Practitioners recommend that anesthesia and analgesia be administered during castration. The objective of this study was to evaluate the effectiveness of bupivacaine liposome suspension, a novel, long-acting, local anesthetic formulation administered as a nerve block at castration. The authors chose to investigate this novel formulation as an alternative to the current industry standards using lidocaine nerve blocks alone or in combination with meloxicam. Thirty male Holstein calves, 16 to 20 wk of age, were enrolled and randomly assigned to one of the four treatment groups prior to surgical castration: 1) bupivacaine liposome suspension block + oral placebo (BUP), 2) lidocaine block + oral placebo (LID), 3) lidocaine block + oral meloxicam (1 mg/kg) (LID + MEL), and 4) saline block + oral placebo (CON). Biomarkers were collected at -24 h and from 0 to 120 h post-castration and included infrared thermography, pressure mat gait analysis, chute defense and behavior scoring (pain and activity), and blood sampling for serum cortisol and prostaglandin E2 metabolites (PGEMs). Responses were analyzed using repeated measures, with calf nested in treatment as a random effect, and treatment, time, and their interaction designated as fixed effects. The results from pressure mat gait analysis show that the CON had a shorter front limb stance time from baseline (-8.73%; 95% confidence interval [CI]: -24.84% to 7.37%) compared with BUP and LID + MEL (>5.70%; 95% CI: -22.91% to 23.79%) (P < 0.03). The CON tended to have an increase in front limb force from baseline (6.31%; 95% CI: -1.79% to 14.41%) compared with BUP, LID, and LID + MEL (<-5.06%; 95% CI: -14.22% to 0.95%) (P < 0.04). The CON displayed higher counts of hunched standing (2.00; 95% CI: 1.68 to 2.32) compared with LID + MEL (1.43; 95% CI: 1.13 to 1.72) (P = 0.05). The CON had higher cortisol concentrations at 24 h (7.70 ng/mL; 95% CI: 1.52 to 13.87 ng/mL) relative to BUP (3.11 ng/mL; 95% CI: -2.56 to 8.79 ng/mL) (P = 0.002). At 4 and 24 h, LID + MEL had lower PGEM concentrations from baseline (-32.42% and -47.84%; 95% CI: -78.45% to -1.80%) compared with CON (27.86% and 47.63%; 95% CI: 7.49% to 82.98%) (P < 0.02). The administration of bupivacaine liposome suspension as a local anesthetic block at the time of castration was as effective at controlling pain as a multimodal approach of lidocaine and meloxicam.

Castration is a routine procedure performed on beef and dairy operations in the United States. All methods of castration cause pain. The American Veterinary Medical Association and the American Association of Bovine Practitioners recommend that anesthesia and analgesia be administered during castration. The objective of this study was to evaluate the effectiveness of bupivacaine liposome suspension, a novel, long-acting, local anesthetic formulation administered as a nerve block at castration, as an alternative to current industry standards using lidocaine nerve blocks alone or in combination with meloxicam. Evidence provided in the current study demonstrates that pain from surgical castration can last up to 120 h post-castration, indicated by changes in ocular temperature, gait analysis, and prostaglandin metabolite concentrations. These data show that the administration of bupivacaine liposome suspension as a local anesthetic block at the time of castration was as effective at controlling pain as a multimodal approach of lidocaine and meloxicam. A single injection that alleviates both perioperative and postoperative pain would be an attractive option for livestock producers to alleviate pain at the time of castration. Further research is needed to discover effective ways of managing pain for extended durations following painful husbandry procedures.

Successful control of Serratia marcescens outbreak in a neonatal unit of a tertiary-care hospital in Spain.

OBJECTIVE: Serratia marcescens is a Gram-negative bacterium that is found in hospital environments and commonly associated with outbreaks in neonatal units. One S. marcescens isolate was detected from a bloodstream culture from a neonate in our hospital that was followed by an outbreak. The aim of this study was to describe the molecular epidemiology of a S. marcescens outbreak in the neonatal unit. METHODS: In order to investigate the outbreak, weekly surveillance rectal swabs were submitted for culture from all patients admitted in this unit from August to September 2018. Environmental samples were obtained from potential sources in September 2018. Typing of isolates was performed by pulsed field gel electrophoresis (PFGE). In addition, we studied the in vitro activity of chlorhexidine against S. marcescens. RESULTS: During this period, 146 infants were hospitalised in our neonatal unit, of which 16 patients had a S. marcescens-positive sample. A total of 36 environmental surveillance samples were collected, and one sample from a stethoscope from an incubator of a colonized baby was positive for S. marcescens. All the 18 isolates, including the isolate from the stethoscope, belonged to a single PFGE cluster. We found that very low concentrations of chlorhexidine, even with application times close to 0 achieved significant reductions in the amount of S. marcescens. CONCLUSION: A unique clone of S. marcescens caused this outbreak, including isolates from patients and from one stethoscope. The outbreak was controlled with the early implementation of specific control measures.

Pacemaker-induced endocarditis by Gordonia bronchialis.

PURPOSE: Gordonia species are known to be opportunistic human pathogens causing secondary infections. We present the second case in the world of endocarditis caused by Gordonia bronchialis and a review of all the cases of endocarditis caused by Gordonia spp. METHODS: The identification was performed by matrix-assisted desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and 16S rRNA gene sequencing were performed to confirm the identification. Antimicrobial susceptibility was performed by MIC test Strip on Mueller-Hinton agar supplemented with 5% defibrinated sheep blood according to Clinical and Laboratory Standards Institute. RESULTS: Pacemaker-induced endocarditis due to Gordonia bronchialis infection was determined in an 88-year old woman. The patient was treated with ceftriaxone and ciprofloxacin until completing 6 weeks from the pacemaker explant with a good evolution. CONCLUSION: The case presented supports the pathogenic role of Gordonia bronchialis as an opportunistic pathogen and highlights the high risk of suffering infections caused by environmental bacteria.

Arthrobacter creatinolyticus: An emerging human pathogen causing urinary tract infection / Arthrobacter creatinolyticus: un patógeno emergente en el ser humano causante de infecciones del tracto urinario

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A combination of photodynamic therapy and antimicrobial compounds to treat skin and mucosal infections: a systematic review.

BACKGROUND: Antimicrobial photodynamic therapy (aPDT) is a growing approach to treat skin and mucosal infections. Despite its effectiveness, investigators have explored whether aPDT can be further combined with antibiotics and antifungal drugs. OBJECTIVE: To systematically assess the in vivo studies on the effectiveness of combinations of aPTD plus antimicrobials in the treatment of cutaneous and mucosal infections. MATERIALS AND METHODS: Searches were performed in four databases (PubMed, EMBASE, Cochrane library databases, ClinicaTrials.gov) until July 2018. The pooled information was evaluated according to the PRISMA guidelines. RESULTS: 11 full-text articles were finally evaluated and included. The best aPDT combinations involved 5-aminolevulinic acid or phenothiazinium dye-based aPDT. In general, the combination shows benefits such as reducing treatment times, lowering drug dosages, decreasing drug toxicity, improving patient compliance and diminishing the risk of developing resistance. The mechanism of action may be that first aPDT damages the microbial cell wall or membrane, which allows better penetration of the antimicrobial drug. LIMITATIONS: The number of studies was low, the protocols used were heterogeneous, and there was a lack of clinical trials. CONCLUSIONS: The additive or synergistic effect of aPDT combined with antimicrobials could be promising to manage skin and mucosal infections, helping to overcome the microbial drug resistance.

Validation and Search of the Ideal Cut-Off of the Sysmex UF-1000i® Flow Cytometer for the Diagnosis of Urinary Tract Infection in a Tertiary Hospital in Spain.

Urinary tract infections (UTI) are one of the most prevalent infections. A rapid and reliable screening method is useful to screen out negative samples. The objective of this study was to validate the Sysmex flow cytometer UF-1000i by evaluating its accuracy, linearity and carry-over; and define an optimal cut-off value to be used in routine practice in our hospital. For the validation of the UF-1000i cytometer, precision, linearity and carry-over were studied in samples with different counts of bacteria, leukocytes and erythrocytes. Between March and June 2016, urine samples were tested in the Clinical Microbiology Laboratory at University Miguel Servet Hospital, in Spain. Samples were analyzed with the Sysmex UF-1000i cytometer, and cultured. Growth of ≥105 CFUs/mL was considered positive. The validation study reveals that the precision in all the variables is acceptable; that there is a good linearity in the dilutions performed, obtaining values almost identical to those theoretically expected; and for the carry-over has practically null values. A total of 1,220 urine specimens were included, of which 213 (17.4%) were culture positive. The optimal cut-off point of the bacteria-leukocyte combination was 138.8 bacteria or 119.8 leukocytes with an S and E of 95.3 and 70.4%, respectively. The UF-1000i cytometer is a valuable method to screen urine samples to effectively rule out UTI and, may contribute to the reduction of unnecessary urine cultures.

Comparing Two Automated Techniques for the Primary Screening-Out of Urine Culture.

Urinary tract infection is the most common human infection with a high morbidity. In primary care and hospital services, conventional urine culture is a key part of infection diagnosis but results take at least 24 h. Therefore, a rapid and reliable screening method is still needed to discard negative samples as quickly as possible and to reduce the laboratory workload. In this aspect, this study aims to compare the diagnostic performance between Sysmex UF-1000i and FUS200 systems in comparison to urine culture as the gold standard. From March to June 2016, 1,220 urine samples collected at the clinical microbiology laboratory of the "Miguel Servet" hospital were studied in parallel with both analysers, and some technical features were evaluated to select the ideal equipment. The most balanced cut-off values taking into account bacteria or leukocyte counts were 138 bacteria/µL or 119.8 leukocyte/µL for the UF-1000i (95.3% SE and 70.4% SP), and 5.7 bacteria/µL or 4.3 leukocyte/µL for the FUS200 (95.8% SE and 44.4% SP). The reduction of cultured plates was 37.4% with the FUS200 and 58.3% with the UF-1000i. This study shows that both techniques improve the workflow in the laboratory, but the UF-1000i has the highest specificity at any sensitivity and the FUS200 needs a shorter processing time.

Mycobacterium tuberculosis genotypes and predominant clones among the multidrug-resistant isolates in Spain 1998-2005.

Although the incidence of tuberculosis (TB) is gradually decreasing in Spain, there is an increase in the proportion of foreign-born cases. This changing scenario is slowly shifting the local TB epidemiology from endemic to imported cases with an increased risk for multidrug-resistant (MDR) and extensively drug resistant (XDR) strains of Mycobacterium tuberculosis complex. MDR/XDR strains from Spain (n=366 MTBC isolates, 1 strain per patient) isolated between 1998 and 2005 were retained for this retrospective analysis. All strains were analyzed by spoligotyping, while 12-loci MIRU-VNTR data were available for 106 isolates from 2003 to 2005. Demographic, phylogenetic, and epidemiologic analyses using anonymized data were collected and analyzed using the SITVIT2 database. Our study provides with a first snapshot of genetic diversity of MDR/XDR-TB in several autonomous regions of Spain. It highlights significantly more of SIT1/Beijing and SIT66/BOV MDR isolates (5.7% and 7.38% respectively) and increasingly more foreign-born cases from Eastern Europe. Future studies should focus on shared genotypes between Spanish and foreign-born patients to decipher the modes of transmission and risk factors involved, and decipher the proportion of imported cases of active disease versus cases of reactivation of latent TB infection among foreign-born individuals.

Superior performance of macroporous over gel type polystyrene as a support for the development of photo-bactericidal materials.

A hexanuclear molybdenum cluster [Mo6I8Ac6]2- (1) has been ionically bound onto macroporous (Pmp) and gel-type (Pgel) resins and their performance as materials for the photodynamic inactivation of microorganisms has been studied. It has been found that 1@Pmp in combination with light is able to reduce 99.999999% of the population of Gram-positive Staphylococcus aureus whereas the activity of 1@Pgel is limited to a 99.99% reduction at the same light dose. The same trend is observed with Gram-negative Pseudomonas aeruginosa. A comprehensive study of both materials has been performed using confocal laser scanning microscopy, thermogravimetric analysis, nitrogen porosimetry, steady state and time resolved fluorometries and diffuse reflectance spectroscopy. The photochemical generation of singlet oxygen (1O2) has been assessed using 9,10-dimethylanthracene as a trap for this reactive oxygen species. It can be concluded that the nature of the polymeric support is of paramount importance for the development of surfaces with bactericidal properties.

In-depth analysis of the genome sequence of a clinical, extensively drug-resistant Mycobacterium bovis strain.

Although human-to-human transmission of Mycobacterium bovis strains and other members of the animal lineage of the tubercle bacilli is a rare event, an extensively drug resistant (XDR) strain, named M. bovis B strain, caused a lethal outbreak in the nineties in Spain. The genome of M. bovis B strain was re-sequenced by SOLiD platform and mapped to the reference M. bovis AF2122/97. The genetic polymorphisms detected have been analysed in depth. One hundred and fifty-eight specific non-synonymous SNPs were detected; ninety-two of these were non-conservative. In addition, one specific 3195-bp insertion could be identified as an ABC transporter gene by homology with tbd2 gene, which was found to be present in other clinical M. bovis strains. Its peculiar phenotype profile of resistance was explained by molecular characteristics, including a 5685-bp specific deletion that revealed a novel polymorphism associated with resistance to paraminosalicilic acid. From a phylogenetical point of view, according to the SNPs detected, M. bovis B could be included into the clonal complex M. bovis European 2. This is the first time that a deep analysis of the whole-genome sequencing of an extensively drug-resistant M. bovis strain is detailed. It offers the explanation for the resistance and found several data to be incorporated for future research.

Klebsiella pneumoniae survives within macrophages by avoiding delivery to lysosomes.

Klebsiella pneumoniae is an important cause of community-acquired and nosocomial pneumonia. Evidence indicates that Klebsiella might be able to persist intracellularly within a vacuolar compartment. This study was designed to investigate the interaction between Klebsiella and macrophages. Engulfment of K. pneumoniae was dependent on host cytoskeleton, cell plasma membrane lipid rafts and the activation of phosphoinositide 3-kinase (PI3K). Microscopy studies revealed that K. pneumoniae resides within a vacuolar compartment, the Klebsiella-containing vacuole (KCV), which traffics within vacuoles associated with the endocytic pathway. In contrast to UV-killed bacteria, the majority of live bacteria did not co-localize with markers of the lysosomal compartment. Our data suggest that K. pneumoniae triggers a programmed cell death in macrophages displaying features of apoptosis. Our efforts to identify the mechanism(s) whereby K. pneumoniae prevents the fusion of the lysosomes to the KCV uncovered the central role of the PI3K-Akt-Rab14 axis to control the phagosome maturation. Our data revealed that the capsule is dispensable for Klebsiella intracellular survival if bacteria were not opsonized. Furthermore, the environment found by Klebsiella within the KCV triggered the down-regulation of the expression of cps. Altogether, this study proves evidence that K. pneumoniae survives killing by macrophages by manipulating phagosome maturation that may contribute to Klebsiella pathogenesis.