Improved culture enrichment broth for isolation of Arcobacter-like species from the marine environment.

Arcobacter-like species are found associated with many matrices, including shellfish in marine environments. The culture media and conditions play a major role in the recovery of new Arcobacter-like species. This study was aimed to develop a culture media for isolation and enhanced growth of Arcobacter-like spp. from marine and shellfish matrices. For this purpose, 14 different Arcobacter-like spp. mostly isolated from shellfish, were grown in 24 different formulations of enrichment broths. The enrichment broths consisted of five main groups based on the organic contents (fresh oyster homogenate, lyophilized oyster either alone or in combination with other standard media), combined with artificial seawater (ASW) or 2.5% NaCl. Optical density (OD420nm) measurements after every 24 h were compared with the growth in control media (Arcobacter broth) in parallel. The mean and standard deviation were calculated for each species in each broth and statistical differences (p < 0.05) among broths were calculated by ANOVA. The results indicated that shellfish-associated Arcobacter-like species growth was significantly higher in Arcobacter broth + 50% ASW and the same media supplemented with lyophilized oysters. This is the first study to have used fresh or lyophilized oyster flesh in the enrichment broth for isolation of shellfish-associated Arcobacter-like spp.

Aliarcobacter vitoriensis sp. nov., isolated from carrot and urban wastewater.

Two isolates, one recovered from a carrot and another one from urban wastewater, were characterized using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences revealed that both isolates clustered together, and were most closely related to Aliarcobacter lanthieri. Multilocus phylogenetic analysis (MLPA) using the concatenated sequences of five housekeeping genes (atpA, gyrA, gyrB, hsp60 and rpoB) suggested that these isolates formed a distinct phylogenetic lineage among the genera derived from the former genus Arcobacter. Whole-genome sequence, in silico DNA-DNA hybridization (isDDH) and the average nucleotide identity (ANI) value between the genome of strain F199T and those of related species confirmed that these isolates represent a novel species. These strains can be differentiated from its phylogenetically closest species A. lanthieri by its inability to growth on 1% glycine and by their enzyme activity of esterase lipase (C8) and acid phosphatase. Our results, by the application of a polyphasic analysis, confirmed that these two isolates represent a novel species of the genus Aliarcobacter, for which the name Aliarcobacter vitoriensis sp. nov. is proposed. The type strain is F199T (=CECT 9230T=LMG 30050T).

Corrigendum (2): Revisiting the Taxonomy of the Genus Arcobacter: Getting Order From the Chaos.

[This corrects the article DOI: 10.3389/fmicb.2018.02077.].

The Use of a DNA-Intercalating Dye for Quantitative Detection of Viable Arcobacter spp. Cells (v-qPCR) in Shellfish.

The genus Arcobacter (Vandamme et al., 1991), comprised of Campylobacter-related species, are considered zoonotic emergent pathogens. The presence of Arcobacter in food products like shellfish, has an elevated incidence worldwide. In this study, we developed a specific viable quantitative PCR (v-qPCR), using the dye propidium monoazide (PMA), for quantification of the viable Arcobacter spp. cells in raw oysters and mussels. The high selectivity of primers was demonstrated by using purified DNA from 38 different species, 20 of them from the genus Arcobacter. The optimization of PMA concentration showed that 20 µM was considered as an optimal concentration that inhibits the signal from dead cells at different concentrations (OD550 from 0.2 to 0.8) and at different ratios of live: dead cells (50:50 and 90:10). The v-qPCR results from shellfish samples were compared with those obtained in parallel using several culture isolation approaches (i.e., direct plating on marine and blood agar and by post-enrichment culturing in both media). The enrichment was performed in parallel in Arcobacter-CAT broth with and without adding NaCl. Additionally, the v-qPCR results were compared to those obtained with traditional quantitative (qPCR). The v-qPCR and the qPCR resulted in c.a. 94% of positive detection of Arcobacter vs. 41% obtained by culture approaches. When examining the reduction effect resulting from the use of v-qPCR, samples pre-enriched in Arcobacter-CAT broth supplemented with 2.5% NaCl showed a higher reduction (3.27 log copies) than that of samples obtained directly and those pre-enriched in Arcobacter-CAT broth isolation (1.05 and 1.04). When the v-qPCR was applied to detect arcobacter from real shellfish samples, 15/17 samples tested positive for viable Arcobacter with 3.41 to 8.70 log copies 1g-1. This study offers a new tool for Arcobacter surveillance in seafood.

Arcobacter lacus sp. nov. and Arcobacter caeni sp. nov., two novel species isolated from reclaimed water.

Two strains (RW43-9T and RW17-10T) recovered from secondary treated wastewater from the Wastewater Treatment Plant (WWTP) in Reus (Spain) were characterized by a polyphasic taxonomic study, showing evidence that they represented two novel Arcobacter species. Based on the 16S rRNA gene for strain RW43-9T, the closest relative was Arcobacter butzleri LMG 10828T (99.9 % similarity), while for strain RW17-10T it was Arcobacter venerupis CECT 7836T (99.4 %). Additionally, multilocus phylogenetic analysis of five concatenated housekeeping genes (atpA, gyrA, gyrB, hsp60 and rpoB) showed that the two strains formed separate branches that are different from known Arcobacter species. Whole genome sequences of the two strains (RW43-9T and RW17-10T) were obtained and they were compared with those of the type strains of their nearest species. Using average nucleotide identity and in silico DNA-DNA hybridization gave values that were below 96 and 70 %, respectively. These results clearly confirm that they represent novel species. Additionally, the phenotypic characterization of the strains allowed their differentiation from other species. Therefore, the strains are proposed as representing two novel species with the names Arcobacter lacus sp. nov. (type strain RW43-9T=CECT 8994T=LMG 29062T) and Arcobacter caeni sp. nov. (type strain RW17-10T=CECT 9140T=LMG 29151T).

Revisiting the Taxonomy of the Genus Arcobacter: Getting Order From the Chaos.

Since the description of the genus Arcobacter in 1991, a total of 27 species have been described, although some species have shown 16S rRNA similarities below 95%, which is the cut-off that usually separates species that belong to different genera. The objective of the present study was to reassess the taxonomy of the genus Arcobacter using information derived from the core genome (286 genes), a Multilocus Sequence Analysis (MLSA) with 13 housekeeping genes, as well as different genomic indexes like Average Nucleotide Identity (ANI), in silico DNA-DNA hybridization (isDDH), Average Amino-acid Identity (AAI), Percentage of Conserved Proteins (POCPs), and Relative Synonymous Codon Usage (RSCU). The study included a total of 39 strains that represent all the 27 species included in the genus Arcobacter together with 13 strains that are potentially new species, and the analysis of 57 genomes. The different phylogenetic analyses showed that the Arcobacter species grouped into four clusters. In addition, A. lekithochrous and the candidatus species 'A. aquaticus' appeared, as did A. nitrofigilis, the type species of the genus, in separate branches. Furthermore, the genomic indices ANI and isDDH not only confirmed that all the species were well-defined, but also the coherence of the clusters. The AAI and POCP values showed intra-cluster ranges above the respective cut-off values of 60% and 50% described for species belonging to the same genus. Phenotypic analysis showed that certain test combinations could allow the differentiation of the four clusters and the three orphan species established by the phylogenetic and genomic analyses. The origin of the strains showed that each of the clusters embraced species recovered from a common or related environment. The results obtained enable the division of the current genus Arcobacter in at least seven different genera, for which the names Arcobacter, Aliiarcobacter gen. nov., Pseudoarcobacter gen. nov., Haloarcobacter gen. nov., Malacobacter gen. nov., Poseidonibacter gen. nov., and Candidate 'Arcomarinus' gen. nov. are proposed.

Do the Escherichia coli European Union shellfish safety standards predict the presence of Arcobacter spp., a potential zoonotic pathogen?

The genus Arcobacter comprises Campylobacter-related species, considered zoonotic emergent pathogens, the presence of which in water has been associated with fecal pollution. Discharges of fecal polluted water into the sea have been considered as one of the main reasons for the presence of Arcobacter in shellfish, and this may represent a risk for public health. In this study, the European Union shellfish food safety criteria based on levels of Escherichia coli were studied in relation to their capacity to predict the presence of Arcobacter species. In addition, the accumulation factor (AF) that measures the concentration ratio between the microbes present in the shellfish and in the water, was also studied for both bacteria. The results show that the presence of E. coli correlated with the presence of the potentially pathogenic species A. butzleri and A. cryaerophilus. However, in 26.1% of the shellfish samples (corresponding to those taken during summer months) E. coli failed to predict the presence of, for instance A. butzleri and A. skirrowii, among other species. In the rest of the samples a significant correlation between the concentration of E. coli and Arcobacter spp. (mussels and oyster; R2=0.744) was found. This study indicates that the presence of E. coli can predict the presence of pathogenic Arcobacter species in shellfish samples harvested from water with temperatures lower than 26.2°C. Consumption of shellfish collected at higher temperatures which may not be permissive to the growth of E. coli but does allow growth of Arcobacter spp., may represent a risk for consumers.

Arcobacter canalis sp. nov., isolated from a water canal contaminated with urban sewage.

Four bacterial strains recovered from shellfish (n=3) and from the water (n=1) of a canal contaminated with urban sewage were recognized as belonging to a novel species of the genus Arcobacter (represented by strain F138-33T) by using a polyphasic characterization. All the new isolates required 2 % NaCl to grow. Phylogenetic analyses based on 16S rRNA gene sequences indicated that all strains clustered together, with the most closely related species being Arcobacter marinus and Arcobactermolluscorum. However, phylogenetic analyses using the concatenated sequences of housekeeping genes (atpA, gyrB, hsp60, gyrA and rpoB) showed that all the novel strains formed a distinct lineage within the genus Arcobacter. Results of in silico DNA-DNA hybridization and the average nucleotide identity between the genome of strain F138-33T and those of the closely related species A. marinus and other relatively closely related species such as A. molluscorum and Arcobacterhalophilus were all below 70 and 96 %, respectively. All the above results, together with the 15 physiological and biochemical tests that could distinguish the newly isolated strains from the closely related species, confirmed that these strains represent a novel species for which the name Arcobacter canalis sp. nov. is proposed, with the type strain F138-33T (=CECT 8984T=LMG 29148T).

Corrigendum: Revisiting the Taxonomy of the Genus Arcobacter: Getting Order From the Caos.

[This corrects the article DOI: 10.3389/fmicb.2018.02077.].

Enhanced recovery of Arcobacter spp. using NaCl in culture media and re-assessment of the traits of Arcobacter marinus and Arcobacter halophilus isolated from marine water and shellfish.

The genus Arcobacter is a relatively poorly known group of bacteria, and the number of new species and sequences from non-culturable strains has increased considerably in recent years. This study investigates whether using media that contain NaCl might help to improve the recovery of Arcobacter spp. from marine environments. To this aim, 62 water and shellfish samples were analysed in parallel, with both a commonly used culture method (enrichment in Arcobacter-CAT broth followed by culture on Blood Agar) and a new one that supplements the Arcobacter-CAT enrichment broth with 2.5% NaCl (w/v) followed by culturing on Marine Agar. The new method yielded ca. 40% more positive samples and provided a higher diversity of known (11 vs. 7) and unknown (7 vs. 2) Arcobacter species. Among the 11 known species recovered, Arcobacter marinus and Arcobacter halophilus were isolated only by this new method. No more strains of these species have been isolated since their original descriptions, both of which were based only on a single strain. In view of that, the phenotypic characteristics of these species are re-evaluated in the present study, using the new strains. Strains of A. halophilus had the same phenotypic profile as the type strain. However, some strains of A. marinus differed from the type strain in that they did not hydrolyse indoxyl-acetate, becoming, therefore, the first Arcobacter species to show a varying ability to hydrolyse indoxyl-acetate. This study shows to what extent a simple variation to the culture media can have a big influence on positive samples and on the community of species recovered.

Characterization of Arcobacter suis isolated from water buffalo (Bubalus bubalis) milk.

During a survey in a dairy plant in Italy, the second strain (strain FG 206) of Arcobacter suis described in the literature was isolated from raw water buffalo milk. The objective of this study was to confirm the species identification, better define the species by comparing its characteristics with those of the reference strain (F41(T) = CECT 7833(T) = LMG 26152(T)) and to investigate its potential clinical relevance by detecting the virulence gene pattern of the new strain. Phenotypical characterization and 16S rRNA-RFLP gave a complete overlap of results for the two strains. As expected, an RFLP pattern common to A. suis and Arcobacter defluvii was obtained by MseI endonuclease digestion, and a pattern specific for A. suis was obtained by BfaI endonuclease digestion. 16S rRNA sequencing and multilocus phylogenetic analysis (MLPA) showed a robust relatedness of strain FG 206 to the A. suis type strain F41(T). The recovery of strain FG 206 from a dairy plant shows that this species of Arcobacter is present in the food chain. Like the type strain recovered from pig meat, the species A. suis may not be confined to a single type of food.

Aplicaciones de la electromiografía de superficie en el deporte / Surface electromyography applications

Las técnicas electrofisiológicas (neurografía y electromiografía de aguja) permiten una aproximación al conocimiento de la función neuromuscular. La electromiografía obtiene la actividad eléctrica del músculo en reposo o activo (contracción voluntaria máxima y estática). En su aplicación clínica, asiste al diagnóstico y al seguimiento de un proceso de tipo neuromuscular. Por otro lado, la electromiografía de superficie (EMGS) o cinesiológica permite estudiar la actividad muscular en acciones dinámicas, siendo aplicable al análisis biomecánico de un gesto, al análisis de la marcha, en estudios de fatiga muscular y de rendimiento deportivo y en áreas como la medicina laboral y la ergonomía. La EMGS ofrece algunas ventajas: es incruenta y permite analizar simultáneamente distintos músculos en movimiento y en acciones de duración ilimitada. El procesado de la señal electromiográfica proporciona parámetros de amplitud y frecuencia para estudios descriptivos y comparativos. No obstante, no permite valorar la musculatura profunda y aporta menos definición que los electromiogramas de aguja(AU)

The electrophysiological techniques (neurography and needle electromyography) allow us an approach to the knowledge of the neuromuscular function. Electromyography obtains the electrical activity from the muscle in rest or in contraction (maximum and static voluntary contraction). In its clinical application, electromyography helps to the diagnosis and follow-up of a process of neuromuscular type. On the other hand, kinesiological or surface electromyography (SEMG) allows the study detecof the muscular activity in dynamics, which we can apply to the biomechanical movement analysis, gait analysis, studies of muscular fatigue, sport performance and applications in work medicine and ergonomics. SEMG brings advantages like the fact that is a bloodless test, of being able to analyze varying muscles at the same time, in motion and in actions of non limited duration. The processed one brings us parameters of amplitude and frequencies, which we will use for descriptive and comparative studies. As a balancing entry, it does not allow us to study deep musculature, and some aspects of definition are lost in the obtained outlines(AU)

Análisis cinescológico de un gesto habitual en enfermería / Kinesiological analysis of an usual gesture in nurses. Comparisonwith an alternative gesture

Objetivos: Estudiar cinemática y cinéticamente un gesto habitual en el personal de enfermería y auxiliar de un centro sociosanitario, para identificar riesgos de lesión lumbar. Estudiar cinemática y cinéticamente un gesto habitual en el personal de enfermería y auxiliar de un centro Sociosantiario, para identificar riesgos de lesión lumbar. Paralelamente, se propone un gesto alternativoy se evalúa si reduce este riesgo de lesión. Métodos: la muestra de voluntarios del personal del Centro en estudio, sin factores de riesgo ni patologías relacionadas con el estudio, es de 27 profesionales (34,9 + 8,2 años de edad; 61,0 + 18,1 Kp de peso, 161,8 + 9,7 cm de altura). Mediante una encuesta al personal se determina el gesto de mayor riesgo subjetivo para este colectivo que será objeto de estudio. Se realiza en laboratorio la grabación cinemática y de actividad muscular de los individuos cuando realizan un traslado de una silla a otra silla de un paciente con hemiplejia derecha. Este gesto se realiza según la forma seguida en el centro y también según una propuesta . Los traslados se desglosan en fase de giro, estabilización y descarga en el destino, ésta última constituye el objeto del estudio. Basado en el modelo biomecánico, cinemáticamente se analiza la postura y el movimiento usado, y cinéticamente se recoge la mediana de la actividad electromiográfica en toda la fase, normalizada a la contracción voluntaria máxima. Paciente y profesional comparan subjetivamente los dos gestos. Resultados: la encuesta de valoración de riesgos indica que es el gesto más escogido con ochenta y ocho respuestas (22,3%) pero con diferencias entre turnos (p=0,02). La sensación del control del enfermo es buena con independencia del método y del gesto utilizados. La postura de contrapeso es la más persistente. En un mismo gesto, ya sea el normal o el alternativo, los auxiliares utilizan diferentes técnicas para transferir al paciente. En ocasiones, en el momento de repetir el gesto, tampoco reproducen de forma exacta la misma técnica o patrón de movimiento. Los cambios introducidos en el ene l gesto alternativo se reflejan en un mayor rango de desplazamiento vertical de S1 y en una mayor inclinación del tronco con respecto a la vertical. No se manifiestan diferencias significativas con respecto a la actividad de los músculos paravertebrales entre los dos gestos propuestos. Conclusiones: La acción considerada de mayor riesgo subjetivo es la reorientación del enfermo hacia otro lugar en una transferencia. La sensación de buen control del paciente es persistente con independencia del gesto utilizado por el auxiliar. Los individuos que practican una postura de contrapeso (comprobada como de bajo riesgo) durante la ejecución del gesto normal persistente en la misma incluso cuando intentan el método alternativo, pero bajan más la posición de la pelvis si comparamos su postura con la del resto de los auxiliares. Este descenso se correlaciona con una disminución de la actividad electromiográfica paravertebral. El gesto alternativo muestra en todos los casos una mayor inclinación de tronco sin un mayor grado de flexión lumbo-pélvico (AU)

Objetives: to define the biomechanical characteristics of a usual gesture in the nursing profession, considered as a risk factor for low back pain. And to compare it with an alternative gesture designed to carry out the same task. Method: Through direct observation and interviewing the personnel, the task and gesture to be studied are selected. A sample of 27 volunteers is gathered for kinematic and muscle activity recording when transferring a patient with right hemiplegia from chair to chair. This gesture is performed as it is “usually practiced” and also in a proposed “alternative”. The movement is divided into turning, stabilizing and unloading phases, the latter being the object of our study. Both patient and professional compare the two gestures subjectively. Results: The risk evaluation survey shows that “reorienting the standing patient towards another place when transferring a patient” is the first choice gesture in 88 responses (22,3%), but with some differences among workshifts (p=0,02). The patient´s control sensation is good, regardless of method and gesture used. The counterweight posture is the most persistent. Within one same gesture, either the usual or alternative gesture, health workers use different techniques to transfer the patient. Those changes introduced in the alternative gesture have resulted in a greater S1 vertical movement range and in a higher trunk tilt in relation to the vertical. Conclusions. The action considered as having higher subjective risk is reorienting the patient towards another place when transferring a patient. People who practice a counterweight posture (defined as low risk) during performance of the usual gesture persist in the same one even when trying the alternative method, lowering the pelvis position if compared with the rest of health workers. This lowering is correlated with a decrease in paraspinal electromyographic activity (lower muscle effort). The correct performance of the alternative method implies greater trunk tilt, allowing the patient to be closer to the health worker, and thus avoiding lumbar flexion, and presenting lower lumbar segment effort (AU)

Pie en la Danza: enfermedades profesionales / The foot in dance: professional diseases

Las vías clínicas son instrumentos de mejora continua de la calidad asistencial, que conjugan la calidad científico-técnica, la calidad gestora y la satisfacción de los pacientes y profesionales para alcanzar los mejores resultados posibles mediante la medicina basada en la evidencia y la eficiencia en los procesos. Objetivos: Analizar los resultados de la vía de septorrinoplastia durante el primer año de la implantación y verificar la repercusión asistencial que ha supuesto su introducción en el servicio de maxilofacial. Metodología: La evaluación de los resultados de la vía se ha obtenido de los documentos de la vía y de la revisión de historias clínicas. La valoración del impacto de la vía clínica se efectuó comparando los resultados de los pacientes intervenidos e incluidos en vía con un grupo control formado por los pacientes intervenidos del mismo proceso durante el año 1999, siendo ambos grupos comparables. Resultados: De los 39 pacientes intervenidos durante el año 2000 de septorrinoplastia en el servicio de maxilofacial 35 fueron incluidos en vía (cobertura del 89,7 por ciento). La estancia media hospitalaria de los pacientes ingresados en vía fue de 2,17 días (DE: 0,62 días), ajustándose a lo programado en vía, frente a la estancia del grupo control de 2,41 días (DE: 1, 24 días).Las variaciones innecesarias debidas a los profesionales fueron un 20 por ciento en el grupo en vía clínica frente al 84,8 por ciento en el grupo control (p < 0,001). Derivadas de estas variaciones, también disminuyeron las variaciones debidas al paciente a un 28,6 por ciento en el grupo en vía, frente a un 56,5 por ciento del grupo control (p = 0,014). Un 97,1 por ciento de los pacientes en vía recibieron la analgesia de forma pautada (el 100 por ciento con pauta correcta) frente al 87 por ciento del grupo control (el 65 por ciento con pauta correcta; p < 0,001).La encuesta de satisfacción con una cobertura del 60 por ciento (21/35) valoró la atención recibida por el servicio con un índice de satisfacción global (IS) del 98,3 por ciento. Conclusiones: La vía clínica de septorrinoplastia, primera vía implantada en el Servicio de Maxilofacial, se revela como un instrumento innovador de la gestión clínica, que permite reducir de manera importante la variabilidad innecesaria, ajustar la estancia hospitalaria, aumentar la efectividad de la asistencia sin menoscabo de la calidad asistencial y conseguir una alta satisfacción del paciente (AU)

Análisis de la marcha mediante el protocolo ELICLINIC / Gait analysis by ELICLINIC protocol

La marcha humana es una actividad muy compleja, tanto por la intervención de múltiples articulaciones y acciones musculares secuenciales en el tiempo, como por el elevado ritmo de variación de algunos de los parámetros que la definen. El uso de sistemas de análisis instrumentados ha supuesto un gran avance en la evaluación y determinación de las posibles anomalías de la misma. La utilización de sistemas computerizados de análisis ofrece una evaluación objetiva y exhaustiva del patrón de locomoción bipodal del ser humano, proporcionando una medida de todas las variables cinemáticas y cinéticas. Esta información es de gran ayuda en el diagnóstico, planificación y control de un proceso rehabilitador. Este artículo aproxima al lector a uno de los sistemas de análisis de la marcha -la aplicación ELICLINIC del sistema ELITE-, en un intento de informar sobre las etapas del estudio, los parámetros que facilita, las aplicaciones y las ventajas e inconvenientes del mismo (AU)