Long-term survive of Aliarcobacter butzleri in two models symbiotic interaction with Acanthamoeba castellanii.

Aliarcobacter butzleri (formerly known as Arcobacter butzleri) is an emerging food-borne zoonotic pathogen that establishes in vitro endosymbiotic relationships with Acanthamoeba castellanii, a free-living amoeba. Previously, we described that this bacterium acts as an endocytobiont of A. castellanii, surviving for at least 10 days in absence of bacterial replication. Thus, the aim of this study was to evaluate the ability of A. butzleri to survive as a long-term endosymbiont of A. castellanii for 30 days in two models of symbiotic interaction with A. castellanii: (i) endosymbiotic culture followed by gentamicin protection assay and (ii) transwell co-culture assay. The results allow us to conclude that A. butzleri is capable of surviving as an endosymbiont of A. castellanii for at least 30 days, without multiplying, under controlled laboratory conditions. In addition, in the absence of nutrients and as both microorganisms remain in the same culture, separated by semi-permeable membranes, A. castellanii does not promote the survival of A. butzleri, nor does it multiply. Our findings suggest that the greater survival capacity of A. butzleri is associated with their endosymbiont status inside A. castellanii, pointing out the complexity of this type of symbiotic relationship.

First case of diarrhea due to Aliarcobacter butzleri (formerly Arcobacter butzleri) in Ecuador.

The first case of acute watery diarrhea disease due to Aliarcobacter butzleri (formerly Arcobacter butzleri) in Ecuador is reported. An infant presented with moderate protein-calorie malnutrition, dehydration and anemia. A curved Gram-negative organism was isolated from stools, having been preliminarily identified by phenotypic characteristics. Definitive identification was achieved by multiplex PCR. Aliarcobacter butzleri was the only pathogenic microorganism isolated. No other entero-pathogens, enterovirus or parasites were found. Our findings strongly suggest that in this specific case, A. butzleri was the etiological agent. Further investigations are needed to develop standardized diagnostic protocols and to establish the prevalence and significance of Aliarcobacter infections in humans.

[Prevention measures to avoid COVID-19 spreading by: from the everyday to the technical-scientific actions.] / Medidas de prevención para evitar el contagio por la COVID-19: de lo cotidiano a lo técnico-científico.

BACKGROUND: In the attempt to reactivate the economy after several months of the pandemic generated by COVID-19, several South American countries have been forced to attenuate the restriction measures created to prevent contagion, leaving all the responsibility to citizens to safeguard their lives. Faced with this situation, the objective of the work was to promote good prevention practices against COVID-19 in the general population to avoid the collapse of the public health system. METHODS: An exploratory review on prevention measures against COVID-19 was carried out in the databases Elsevier, Pubmed, Wiley, Scopus, Scielo and, in general, in academic Google, to obtain gray information. RESULTS: The role of seven individual protection and prevention measures and barriers to stop the advance of community infections by COVID-19 were described and analyzed. Studies showed that the use of preventive measures can play a critical role in containing the pandemic. However, they can also provide a false security of protection, predisposing to neglect the correct use and handling of all containment measures. CONCLUSIONS: Given the drastic rise in infections and deaths from COVID-19 in Latin America, it becomes necessary to continue with biosecurity campaigns so that citizens are clear that prevention is an integrated process and that the use of one measure does not replace another.

OBJETIVO: En el intento de reactivar la economía tras varios meses de pandemia generada por COVID-19, varios países sudamericanos se han visto obligados a reducir las medidas de restricción creadas para evitar contagios, dejando toda la responsabilidad a la ciudadanía a salvaguardar sus vidas. Ante esta situación, el objetivo del trabajo fue promover buenas prácticas de prevención contra COVID-19 en la población general para evitar el colapso del sistema público de salud. METODOS: Se realizó una revisión exploratoria sobre medidas de prevención contra COVID-19 en las bases de datos Elsevier, Pubmed, Wiley, Scopus, Scielo y, en forma general, en Google académico para obtener información gris. RESULTADOS: Se describieron y analizaron el rol de siete medidas y barreras de protección y prevención individual para detener el avance de los contagios comunitarios por COVID-19. Los estudios mostraron que el uso de medidas de prevención puede desempeñar un papel fundamental en la contención la pandemia. No obstante, también pueden brindan una falsa seguridad de protección, predisponiendo a dejar de lado el uso y manejo correcto de todas las medidas de contención. CONCLUSIONES: Ante el ascenso drástico de los contagios y muerte por la COVID-19 en América Latina, se vuelve necesario continuar con las campañas de bioseguridad para que la ciudadanía tenga claro que la prevención es un proceso integrado y que el uso de una medida no reemplaza otra.

Optimal ECG Lead System for Exercise Assessment of Ischemic Heart Disease.

The diagnostic value of an ECG exercise test in diagnosis of ischemic heart disease (IHD) is limited. We investigated whether it is possible to develop a method for diagnosis of IHD which uses a low number of optimal ECG leads and has a higher diagnostic efficiency than conventional exercise ECG. This study was carried out on 43 patients. The 67-lead high-resolution ECG was recorded at rest and during exercise. The diagnostic value of ST segment depression (ΔST60) and T-wave morphology change (δT) determined in optimized ECG lead configurations was higher than for the standard 12-lead ECG. The best results were obtained for δT determined from 6 ECG electrode locations where sensitivity and specificity were 70% and 69% whereas for the standard exercise ECG were 63% and 62%, respectively. The small number of ECG leads used allows for easy hardware implementation of the methods for use in clinical settings.

Transcriptional analysis of flagellar and putative virulence genes of Arcobacter butzleri as an endocytobiont of Acanthamoeba castellanii.

Arcobacter butzleri is an emerging foodborne zoonotic pathogen that has been isolated from environmental water sources. This pathogen establishes in vitro endosymbiotic relationships with Acanthamoeba castellanii, a free-living amoeba found in environmental matrices such as soil and water. The principal aim of this study was to analyse the transcriptional pattern of flagellar (flaA-flaB-flgH-motA) and other putative virulence genes (ciaB-cadF-mviN-pldA) of A. butzleri during its interaction with A. castellanii by quantitative real-time PCR. The transcriptional analysis showed up-regulation of all genes analysed before A. butzleri became established as an endocytobiont of A. castellanii. In contrast, while A. butzleri remains an endocytobiont, a significant and sustained decrease in the transcription of all analysed genes was observed. Our findings suggest that A. butzleri requires a biphasic transcriptional pattern of flagellar and other putative virulence genes to establish an endosymbiotic relationship with A. castellanii.

Campylobacter species in chilean patients with chronic periodontitis / Espécies de Campylobacter em pacientes chilenos com periodontite crônica

The purpose of this study is to identify the presence of Campylobacter species (C. rectus, C. concisus, C. gracilis and C. ureolyticus) in Chilean patients with chronic periodontitis, and to establish the relationship of these microorganisms with the periodontal conditions of smoker and non-smoker patients. Subgingival plaque samples were collected from four periodontal-affected tissue sites of 15 smoker and 15 non-moker patients with chronic periodontitis. A sample per quadrant was obtained, with a probing depth of 6 mm and an insertion loss of >3 mm in each patient. Polymerase chain reaction (PCR) was performed with the specific 16S rDNA primers for the molecular detection of C. rectus and C. gracilis, specific cpn60 primers for C. concisus and hsp60 primers gene for C. ureolyticus. Campylobacter species showed an overall prevalence of 93.3% in periodontal patients, while C. rectus was the most frequent one (80%), followed by C. concisus (66.7%), C. gracilis (33.3%), and C. ureolyticus (10%). Only C. gracilis showed a statistically significant (p = 0.002) association with chronic periodontitis among samples from smoker and nonsmoker patients. A high prevalence of the Campylobacter genus in the analyzed populations (93.3%) was found, being C. rectus the most frequent (80%) species. Besides, C. gracilis showed a statistically significant association between smoker state and chronic periodontitis.

O objetivo deste estudo é identificar a presença de espécies de Campylobacter (C. rectus, C. concisus, C. gracilis e C. ureolyticus) em pacientes chilenos com periodontite crônica, e estabelecer a relação desses microrganismos com as condições periodontais de pacientes fumantes e não fumantes. Amostras de placa subgengival foram coletadas de quatro locais de tecido periodontal afetado de 15 pacientes fumantes e 15 não fumantes com periodontite crônica. Obteve-se uma amostra por quadrante, com profundidade de sondagem 6 mm e perda de inserção > 3 mm em cada paciente. A reação em cadeia da polimerase (PCR) foi realizada com os primers específicos 16S rDNA para a detecção molecular de C. rectus e C. gracilis, primers específicos de cpn60 para C. concisus e gene primers de hsp60 para C. ureolyticus. As espécies de Campylobacter apresentaram uma prevalência geral de 93,3% nos pacientes periodontais, enquanto C. rectus foi o mais frequente (80%), seguido por C. concisus (66,7%), C. gracilis (33,3%) e C. ureolyticus (10%). Apenas C. gracilis apresentou associação estatisticamente significante (p = 0,002) com a periodontite crônica entre as amostras de pacientes fumantes e não fumantes. Observou-se alta prevalência do gênero Campylobacter nas populações analisadas (93,3%), sendo C. reto as espécies mais frequentes (80%). Além disso, C. gracilis mostrou associação estatisticamente significante entre estado de fumante e periodontite crônica.

[Prevalence and antimicrobial behavior of Campylobacter jejuni and Campylobacter coli in children with diarrhea in Loja city, Ecuador]. / Prevalencia y caracterización del perfil de susceptibilidad antimicrobiana de Campylobacter jejuni y Campylobacter coli en niños con diarrea de la ciudad de Loja, Ecuador.

INTRODUCTION: Campylobacter is an important agent of diarrhea in humans. In Ecuador, the information on Campylobacter is scarce and there are not antecedents about antimicrobial susceptibility. OBJECTIVE: To describe Campylobacter prevalence in children with diarrhea and their behavior against five antimicrobials in vitro. METHOD: We studied 253 children with diarrhea aging 7 months to 9 years who consulted for diarrhea in two hospitals in the city of Loja. Fecal samples were cultured and identification by tests by phenotypic tests and multiplex PCR. Susceptibility to 5 antibiotics was determined by the disc-diffusion method. RESULTS: Campylobacter was found in 16 (6.3%) children, being C. jejuni the most frequent one (5.1%), followed by C. coli (1.2%). All strains were susceptible to gentamicin and ampicillin/clavulanic acid, being found low resistance to ampicillin and erythromycin and high resistance to ciprofloxacin.

Prevalencia y caracterización del perfil de susceptibilidad antimicrobiana de Campylobacter jejuni y Campylobacter coli en niños con diarrea de la ciudad de Loja, Ecuador / Prevalence and antimicrobial behavior of Campylobacter jejuni and Campylobacter coli in children with diarrhea in Loja city, Ecuador

Resumen Introducción: Campylobacter es un importante agente de diarrea en el ser humano. En Ecuador, la información sobre Campylobacter es escasa y no existen antecedentes de susceptibilidad antimicrobiana. Objetivo: Describir la prevalencia de Campylobacter en niños con diarrea y su comportamiento in vitro frente a cinco antimicrobianos. Método: Se estudiaron 253 niños entre siete meses y 9 años de edad, que consultaron por diarrea en dos hospitales de la ciudad de Loja. Se realizó cultivo de muestras fecales e identificación por pruebas fenotípicas y por RPC múltiple. La susceptibilidad antimicrobiana fue determinada por el método de difusión en disco. Resultados: Campylobacter fue diagnosticado en 16 (6,3%) de las muestras, aislándose C. jejuni en 13 (5,1%) y C. coli en 3 (1,2%). Todas las cepas fueron susceptibles a gentamicina y ampicilina/ ácido clavulánico, con baja resistencia a ampicilina y eritromicina y alta resistencia a ciprofloxacina.

Introduction: Campylobacter is an important agent of diarrhea in humans. In Ecuador, the information on Campylobacter is scarce and there are not antecedents about antimicrobial susceptibility. Objective: To describe Campylobacter prevalence in children with diarrhea and their behavior against five antimicrobials in vitro. Method: We studied 253 children with diarrhea aging 7 months to 9 years who consulted for diarrhea in two hospitals in the city of Loja. Fecal samples were cultured and identification by tests by phenotypic tests and multiplex PCR. Susceptibility to 5 antibiotics was determined by the disc-diffusion method. Results: Campylobacter was found in 16 (6.3%) children, being C. jejuni the most frequent one (5.1%), followed by C. coli (1.2%). All strains were susceptible to gentamicin and ampicillin/clavulanic acid, being found low resistance to ampicillin and erythromycin and high resistance to ciprofloxacin.

Occurrence and antimicrobial resistance of Campylobacter jejuni and Campylobacter coli isolated from domestic animals from Southern Ecuador / Ocorrência e comportamento antimicrobiano de espécies termotolerantes de Campylobacter isoladas de animais domésticos no sul do Equador

ABSTRACT: Occurrence and antimicrobial resistance profiles of Campylobacter jejuni and C. coli strains isolated from fecal samples of dogs, pigs, cows and hens in southern Ecuador were studied. Of the 250 samples studied, 84 (33.6%) were Campylobacter positive, with C. jejuni being more frequent (78.6%) than C. coli (21.4%), with the exception of porcine samples, from which C. coli was the only species isolated. Multidrug resistance was reported in 10 Campylobacter strains (11.9%), four C. jejuni and six C. coli, and was always associated with nalidixic acid and ciprofloxacin resistance. All of the amoxicillin-resistant strains were susceptible to amoxicillin-clavulanic acid and were therefore beta-lactamase producers. However, one strain of C. jejuni remained resistant with additional resistance to gentamycin. This is an uncommon resistance pattern in Campylobacter and could reflect different resistance mechanisms.

RESUMO: A ocorrência e resistência antimicrobiana de cepas de Campylobacter jejuni e C. coli isoladas de amostras fecais obtidas de cães, suínos, vacas e galinhas no sul do Equador foram determinadas. Das 250 amostras estudadas, 84 (33.6%) foram Campylobacter positivas, sendo C. jejuni mais frequente (78.6%) que C. coli (21.4%) com a exceção das amostras de suínos, das quais só fora isolada a espécie C. coli. Multirresistência foi encontrada em 10 cepas (11,9%), quatro C. jejuni e 6 C. coli, sempre associada à resistência ao ácido nalidíxico e à ciprofloxacina. Todas as cepas resistentes à ampicilina foram susceptíveis para amoxicilina-ácido clavulánico demonstrando que foram produtoras de beta-lactamase. Porém, uma cepa de C. jejuni permaneceu sendo resistente, apresentando, também, resistência à gentamicina. Este é um padrão de multirresistência incomum em Campylobacter e pode refletir diferentes mecanismos de resistência.

Occurrence and antimicrobial susceptibility of thermophilic Campylobacter species isolated from healthy children attending municipal care centers in Southern Ecuador.

The prevalence and antimicrobial susceptibility of Campylobacter jejuni and C. coli strains in healthy, well-nourished children of middle socioeconomic level from Southern Ecuador were determined. Among the 127 children studied, 17 (13.4%) harbored Campylobacter sp. corresponding to C. jejuni (7.1%) and C. coli (6.3%) with a higher concentration of C. jejuni among boys (8.6%) and C. coli (8.8%) among girls. C. jejuni showed high resistance to nalidixic acid and ciprofloxacin (77.8%), but susceptibility to all other antimicrobials tested. C. coli strains showed resistance to more antibiotics than C. jejuni strains including resistance to nalidixic acid (75%), ciprofloxacin (75%), erythromycin (12.5%) and ampicillin (28.6), but susceptible to gentamicin and amoxicillin/clavulanic acid.

Intrinsic and Extrinsic Aspects on Campylobacter jejuni Biofilms.

Biofilm represents a way of life that allows greater survival of microorganisms in hostile habitats. Campylobacter jejuni is able to form biofilms in vitro and on surfaces at several points in the poultry production chain. Genetic determinants related to their formation are expressed differently between strains and external conditions are decisive in this respect. Our approach combines phylogenetic analysis and the presence of seven specific genes linked to biofilm formation in association with traditional microbiology techniques, using Mueller Hinton and chicken juice as substrates in order to quantify, classify, determine the composition and morphology of the biomass of simple and mixed biofilms of 30 C. jejuni strains. It also evaluates the inhibition of its formation by biocides commonly used in industry and also by zinc oxide nanoparticles. Genetic analysis showed high heterogeneity with the identification of 23 pulsotypes. Despite the diversity, the presence of flaA, cadF, luxS, dnaJ, htrA, cbrA, and sodB genes in all strains shows the high potential for biofilm formation. This ability was only expressed in chicken juice, where they presented phenotype of a strong biofilm producer, with a mean count of 7.37 log CFU/mL and an ultrastructure characteristic of mature biofilm. The composition of simple and mixed biofilms was predominantly composed by proteins. The exceptions were found in mixed biofilms with Pseudomonas aeruginosa, which includes a carbohydrate-rich matrix, lower ability to sessile form in chicken juice and compact architecture of the biofilm, this aspects are intrinsic to this species. Hypochlorite, chlorhexidine, and peracetic acid were more effective in controlling viable cells of C. jejuni in biofilm, but the existence of tolerant strains indicates exposure to sublethal concentrations and development of adaptation mechanisms. This study shows that in chicken juice C. jejuni presents greater potential in producing mature biofilms.

Biotyping of Arcobacter butzleri isolated from poultry products: rescuing a phenotypic method / Biotipagem de Arcobacter butzleri desde produtos aviares: resgatando um método fenotípico

Arcobacter butzleri is an emergent zoonotic foodborne pathogen associated to enteritis and occasionally to bacteremia in human beings. Biotyping of this bacterium is important in order to establish the circulating strains and its dissemination routes. The purpose of this work was to determine the circulating A. butzleri biotypes in poultry products for human consumption in Southern Chile using the method proposed by Lior and Woodward, in order to explore the possibility of introducing this biotyping scheme as a routine laboratory tool. From the 60 strains studied the prevalent biotypes were 8A, 8B, 7A, 4A and 4B. The most frequently isolated biotype, independently of the sample of origin, was 8A with (44 strains, 73.3%). The less frequently isolated biotype was 4B (two strains 3.3%). The biotyping method used results to be simple, easy to handle and yields stable results. Therefore, it might be rescued to be used as a phenotypic tool for epidemiological marking of A. butzleri.

Arcobacter butzleri é um patógeno emergente, zoonótico e de transmissão alimentar, associado a enterite e, ocasionalmente, a bacteremia em seres humanos. A biotipagem desta bactéria é importante para estabelecer os biótipos circulantes e suas rotas de disseminação. Os objetivos deste trabalho foram determinar os biótipos de A. butzleri circulantes em alimentos de origem aviar para consumo humano, no sul do Chile, explorando a possibilidade de introduzir o método de biotipagem proposto por Lior e Woodward como uma ferramenta de rotina no laboratório. Entre as 60 cepas estudadas, os biótipos 8A, 8B, 7A e 4B foram os mais prevalentes. O biótipo mais frequentemente isolado, independentemente da amostra de origem, foi o biótipo 8A (44 cepas, 73,3%). O biótipo 4B apresentou a menor frequência de isolamento (duas cepas, 3,3%). O método de biotipagem utilizado resultou ser simples de executar, fácil de manusear e produz resultados estáveis. Portanto, pode ser resgatado para ser usado como uma ferramenta fenotípica para marcação epidemiológica de A. butzleri.

World Health Organization Ranking of Antimicrobials According to Their Importance in Human Medicine: A Critical Step for Developing Risk Management Strategies to Control Antimicrobial Resistance From Food Animal Production.

Antimicrobial use in food animals selects for antimicrobial resistance in bacteria, which can spread to people. Reducing use of antimicrobials-particularly those deemed to be critically important for human medicine-in food production animals continues to be an important step for preserving the benefits of these antimicrobials for people. The World Health Organization ranking of antimicrobials according to their relative importance in human medicine was recently updated. Antimicrobials considered the highest priority among the critically important antimicrobials were quinolones, third- and fourth-generation cephalosporins, macrolides and ketolides, and glycopeptides. The updated ranking allows stakeholders in the agriculture sector and regulatory agencies to focus risk management efforts on drugs used in food animals that are the most important to human medicine. In particular, the current large-scale use of fluoroquinolones, macrolides, and third-generation cephalosporins and any potential use of glycopeptides and carbapenems need to be addressed urgently.

Arcobacter butzleri survives within trophozoite of Acanthamoeba castellanii.

The survival of three Arcobacter butzleri strains inside Acanthamoeba castellanii was assessed using axenic cultures of A. castellanii that were inoculated with the tested strains and incubated at 26°C under aerobic conditions for 240h. The behavior of bacteria in contact with amoebae was monitored using phase contrast microscopy. The bacterial survival rate within amoebae was assessed through counting colony forming units, using the gentamicin protection assay. All A. butzleri strains were able to survive during 240h within the amoebae, thus suggesting that (i) A. butzleri resists the amoebic digestion processes at least for the analyzed time; (ii) that A. castellanii could serve as an environmental reservoir for this bacterium, probably acting as a transmission vehicle for A. butzleri.

[In vitro antibacterial activity of Chilean propolis against Helicobacter pylori]. / Actividad antibacteriana in vitro de propóleos chilenos sobre Helicobacter pylori.

INTRODUCTION: Propolis is a natural product derived from beekeeping. It has anesthetic, anti-inflammatory, immune-stimulant and antibacterial properties on grampositive and gramnegative bacteria. However, little is known regarding its activity on Helicobacter pylori. This bacteria colonizes about half of the world's population and is associated with chronic gastritis, peptic ulcer and gastric cancer. OBJECTIVE: The aim of this study was to evaluate the inhibitory activity of 22 propolis extracts from nine of the 11 beekeeping Chilean regions on 10 strains of H. pylori isolated from gastric mucosa. METHODS: The antibacterial activity of the extracts was determined using the well diffusion method and diffusion disks. RESULTS: 100% of the extracts were active on the tested strains, showing inhibition halos equal to or greater than 15 mm by both methods. CONCLUSIONS: Our results show an effective anti H. pylori activity of propolis. However, additional microbiological studies are needed before a potential clinical utility of these natural products is warranted.

Actividad antibacteriana in vitro de propóleos chilenos sobre Helicobacter pylori / In vitro antibacterial activity of Chilean propolis against Helicobacter pylori

Introduction: Propolis is a natural product derived from beekeeping. It has anesthetic, anti-inflammatory, immune-stimulant and antibacterial properties on grampositive and gramnegative bacteria. However, little is known regarding its activity on Helicobacter pylori. This bacteria colonizes about half of the world’s population and is associated with chronic gastritis, peptic ulcer and gastric cancer. Objective: The aim of this study was to evaluate the inhibitory activity of 22 propolis extracts from nine of the 11 beekeeping Chilean regions on 10 strains of H. pylori isolated from gastric mucosa. Methods: The antibacterial activity of the extracts was determined using the well diffusion method and diffusion disks. Results: 100% of the extracts were active on the tested strains, showing inhibition halos equal to or greater than 15 mm by both methods. Conclusions: our results show an effective anti H. pylori activity of propolis. However, additional microbiological studies are needed before a potential clinical utility of these natural products is warranted.

Introducción: El propóleos es un producto natural derivado de la apicultura que tiene propiedades anestésicas, anti-inflamatorias, inmuno-estimulantes y antibacterianas. Ejerce su acción sobre distintas bacterias grampositivas y gramnegativas. Sin embargo, es muy poco lo que se sabe en relación a su actividad sobre H. pylori, bacteria asociada con gastritis crónica, úlcera gastro-duodenal y cáncer gástrico y que coloniza a alrededor de la mitad de la población mundial. Objetivo: Evaluar la actividad inhibitoria de 22 extractos de propóleos de orígenes botánicos diferentes, provenientes de nueve de las once zonas mielíferas de Chile, en la época de otoño, sobre 10 cepas de H. pylori aisladas de mucosa gástrica. Metodología: La actividad antibacteriana de los extractos se determinó a través del método de difusión en pocillos y de difusión en discos. Resultados: 100% de los extractos fueron activos sobre las cepas ensayadas, observándose halos de inhibición iguales o mayores a 15 mm en ambos métodos. Conclusiones: Los datos obtenidos in vitro en el presente estudio muestran una efectiva actividad anti H. pylori de los propóleos chilenos, siendo necesario estudios microbiológicos y farmacológicos adicionales para avanzar en una posible utilidad clínica de estos productos naturales.