Diferencia en la expresión del ligando de receptor activador para el factor nuclear K B, en tejido tumoral frente a la infección por diferentes patógenos periodontales / Difference in receptor activator of nuclear factor K B expression in tumoral tissue by different periodontal pathogens

La enfermedad periodontal es una de las principales causas de pérdida dentaria. Clínicamente, esta patología, mediada por la desregulación del sistema inmune producto de una disbiosis ocurrida en el surco gingival, inicia con la inflamación de la encía y evoluciona con el daño irreversible de los tejidos que rodean el diente. El hueso alveolar es uno de los tejidos afectados esta patología, esto debido a la activación de osteoclastos por la sobreexpresión de la proteína RANKL en el huésped. El propósito de este trabajo es determinar el nivel de sobreexpresión de RANKL, en un modelo de células tumorales U2OS, frente a la infección con Porphyromonas gingivalis y Prevotella intermedia. Para identificar el nivel de RANKL, se definieron cuatro grupos: Un grupo control, no tratado; Grupo PG, tratado con P. gingivalis; Grupo PI, tratado con P. Intermedia; y un grupo PG+PI, tratado con ambas bacterias. El nivel relativo de la proteína RANKL fue determinado en el sobrenadante y en los extractos celulares de manera independiente, mediante la técnica Western blot. En sobrenadantes, el grupo PG mostró mayores niveles de RANKL comparados con PI (p < 0,05). En extractos celulares los niveles fueron mayores en el grupo PG+PI (p < 0,05). El grupo PI mostró los niveles más bajos de RANKL. La infección polimicrobiana resulta en una mayor expresión de RANKL en células tumorales U2OS, mientras que frente a la infección P. gingivalis, se observó mayor cantidad de RANKL soluble.

SUMMARY: Periodontal disease is one of the main causes of tooth loss. Clinically, this pathology, mediated by the deregulation of the immune system due to a dysbiosis occurred in the gingival sulcus, begins with the inflammation of the gum and evolves with the irreversible damage of the tissues that surround the tooth. Alveolar bone is one of the most affected tissues by this disease, due to the activation of osteoclasts by the upregulation of RANKL in the host. The aim of this study is to determine the increase of RANKL, in a U2OS tumor cells model, inoculated with Porphyromonas gingivalis and Prevotella intermedia. To identify the level of RANKL, four groups were defined: A control group, not treated; PG group, treated with P.gingivalis; PI group, treated with P. intermedia; and a PG+PI group, treated with both bacteria. The relative level of RANKL was determined in the supernatant and cell extracts independently, using the Western blot technique. In supernatants, the PG group showed higher RANKL levels compared to PI (p < 0.05). In cell extracts the levels were higher in the PG+PI group (p < 0.05.). The PI group showed the lowest levels of RANKL.Polymicrobial infection results in a greater expression of of soluble RANKL was observed.

Actividad antimicrobiana in vitro del extracto de hojas de Bixa orellana L. contra bacterias anaerobias asociadas a la vaginosis bacteriana y Lactobacillus spp / In vitro antimicrobial activity of Bixa orellana L. leaves extract against anaerobic bacteria associated to bacterial vaginosis and Lactobacillus spp.

Objetivos . Describir la actividad antimicrobiana in vitro del extracto metanólico de las hojas de Bixa orellana L. contra las bacterias anaerobias asociadas a la vaginosis bacteriana y Lactobacillus spp. Materiales y métodos . Se incluyeron en el estudio ocho cepas de referencia ATCC; Gardnerella vaginalis, Prevotella bivia, Peptococcus niger, Peptostreptococcus anaerobius, Mobiluncus curtisii, Atopobium vaginae, Veillonella parvula y Lactobacillus crispatus, y 22 aislamientos clínicos; once aislados de Gardnerella vaginalis y once aislados de Lactobacillus. La susceptibilidad antimicrobiana se determinó mediante el método de difusión en agar. La concentración mínima inhibitoria (CMI) y la concentración bactericida mínima (CBM) fueron determinadas utilizando el método de dilución en agar y un método de dilución modificado, respectivamente. Resultados . Todas las cepas de referencia ATCC tuvieron un alto nivel de susceptibilidad al extracto, con excepción de P. vibia, V. parvula y L. crispatus. Interesantemente, los aislamientos clínicos de G. vaginalis y la cepa ATCC de G. vaginalis fueron los más susceptibles al extracto dados los bajos valores de CMI (1,0 - 2,0 mg/mL) y CBM (1,0 - 4,0 mg/mL), mientras que, los aislamientos clínicos de Lactobacillus spp. y la cepa ATCC de L. crispatus fueron los menos susceptibles debido a los altos valores de CMI (32,0 mg/mL) y CBM (≥ 32,0 mg/mL). Conclusiones . Los experimentos in vitro sugieren que el extracto posee propiedades antibacterianas selectivas dada su alta actividad contra bacterias anaerobias asociadas a vaginosis bacteriana y baja actividad contra especies de Lactobacillus.

Objective. To describe the in vitro antimicrobial activity of the methanolic extract of Bixa orellana L. leaves against anaerobic bacteria associated to bacterial vaginosis and Lactobacillus spp. Materials and methods. Eight ATCC reference strains; Gardnerella vaginalis, Prevotella bivia, Peptococcus niger, Peptostreptococcus anaerobius, Mobiluncus curtisii, Atopobium vaginae, Veillonella parvula, and Lactobacillus crispatus, and twenty-two clinical isolates; eleven Gardnerella vaginalis and eleven Lactobacillus strains, were included in the study. The antimicrobial susceptibility was determined by the agar diffusion method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined by using agar dilution and a modified dilution plating method, respectively. Results. All ATCC reference strains showed high levels of susceptibility to the extract, except P. vibia, V. parvula and L. crispatus. Interestingly, all G. vaginalis clinical isolates and the G. vaginalis ATTC strain were the most susceptible to the extract, given their low MIC (1.0 - 2.0 mg/mL) and MBC (1.0 - 4.0 mg/mL) values, whereas, the Lactobacillus spp. clinical isolates and the L. crispatus ATCC strain were the least susceptible bacteria given their high MIC (32.0 mg/mL) and MBC (≥ 32.0 mg/mL) values. Conclusions. In vitro experiments suggest that the extract possesses selective antimicrobial properties given its high activity against bacterial vaginosis-associated anaerobic bacteria and low activity against Lactobacillus species.

Diagnóstico microbiológico de infección por Clostridioides difficile en épocas de COVID-19 en un hospital pediátrico de alta complejidad / Microbiological diagnosis of Clostridioides difficile infection in times of COVID-19 in a high complexity pediatric hospital

Resumen La infección por Clostridioides difficile (ICD) puede variar desde diarrea hasta megacolon tóxico. Los objetivos del trabajo fueron mostrar la variación en el número de casos diagnosticados de ICD en este laboratorio entre 2020, cuando comenzó la pandemia de COVID-19 y 2019 y 2021 y detallar los casos precedidos por la infección de SARS-CoV-2. El presente es un estudio retrospectivo observacional en el que se registraron el número total de muestras procesadas con sospecha de ICD y el de positivas y los antecedentes clínicos de pacientes con ICD hasta dos meses después de su diagnóstico de COVID-19. Durante 2020 se procesaron menos muestras que en 2019 y 2021; sin embargo, el porcentaje de positividad fue de 13,1%, 7,2% y 7,8%, respectivamente. Esto pudo deberse a mejoras en el criterio clínico al momento de seleccionar las muestras con sospecha de ICD.

Abstract Clostridioides difficile infection (CDI) can cause anything from diarrhea to toxic megacolon. The objectives of this study were: to show the variation in the number of diagnosed cases of CDI in this center, comparing 2020, when the COVID-19 pandemic began, with 2019 and 2021 and to detail cases preceded by SARS-CoV-2 infection. This is an observational retrospective study in which the total number of samples processed with suspected CDI were recorded. The positive ones and the clinical history of patients with a diagnosis of CDI up to two months after their diagnosis of SARS-CoV-2 infection were recorded as well. During 2020 a smaller number of samples were processed. However, during this year the percentage of positivity was 13.1% vs. 7,2% and 7.8% during 2019 and 2021, respectively. It is believed that this may have been due to improvements in clinical suspicion and sample selection for CDI diagnosis.

Resumo A infecção por Clostridioides difficile (ICD) pode causar desde diarreia até megacólon tóxico. Os objetivos desta apresentação foram: mostrar a variação do número de casos diagnosticados de ICD neste laboratório, entre 2020 quando começou a pandemia de COVID-19 e 2019 e 2021 e, detalhar os casos precedidos pela infecção por SARS-CoV-2. Esse estudo foi retrospectivo observacional e foram registrados: o número total de amostras processadas com suspeita de ICD e de amostras positivas e os antecedentes clínicos daqueles pacientes com diagnóstico de ICD até dois meses após o diagnóstico de COVID 19. Durante 2020, foram processadas menos amostras do que em 2019 e 2021; no entanto, o percentual de positividade foi de 13,1%, 7,2% e 7,8%, respectivamente. Isso pode ter sido resultado de melhorias no critério clínico na hora de selecionar as amostras com suspeita de ICD.

Diversity of aerobic and facultative anaerobic bacteria forming the supragingival biofilm in healthy children

Aims@#The aim of this study was to reveal the aerobic and facultative anaerobic bacterial diversity in the supragingival biofilms of healthy caries-free children.@*Methodology and results@#Biofilm sampling was performed by a specialist in the morning at least 12 h after tooth brushing from 6-10 years old children who came to Suleyman Demirel University Pedodontics Clinic. Samples were cultured on selected media. Purified isolates were identified according to 16S rRNA gene sequences. Totally 180 bacterial isolates had obtained. These isolates were identified as 36 different bacterial species belonging to Firmicutes, Proteobacteria and Actinobacteria. Species with the highest number of isolates were Lactobacillus and Staphylococcus.@*Conclusion, significance and impact of study@#In this study, a total of 180 bacterial isolates had obtained. These isolates were identified as 36 different bacterial species belonging to 3 bacterial phyla (Firmicutes, Proteobacteria and Actinobacteria) and 9 bacterial genera. Although our study has certain limitations because it is dependent on culture, since there is no study previously conducted in Turkey to our knowledge, it contains important findings. The research findings reported in this paper will serve as the foundation for studies to be conducted to understand the oral colonization and biofilm dynamics of children and to take protective measures accordingly.

Anal abscess microbiology as an anal fistula predictor / Microbiologia de abscesso anal como preditivo de fístula anal

ABSTRACT In order to determine the microbiology of anal abscess as a predictor of anal fistulas in patients who attended the external consultation of the Coloproctology unit of the Dr. Antonio María Pineda University Central Hospital, a prospective, descriptive, longitudinal study was conducted from September 2018 to July 2019. In this study, the population consisted of patients with a diagnosis of anal abscess, without associated comorbidities or contraindications for surgery, who agreed to be included in the study. A non-probabilistic, intentional sample consisting of 42 patients was determined. An appointment-based study protocol was applied by outpatient for patients who met the inclusion criteria applied, to perform due medical history through anamnesis, physical examination and culture taking of suppuration from the anal abscess to subsequently establish medical and surgical behavior thereof. The results were expressed in absolute numbers and percentages, a prevalence of ischiorectal abscesses was observed, followed by deep post-anal space abscesses. Anaerobic bacteria were isolated in 100% of the sample. In all fistulized patients, E. Coli was isolated as a predominant germ.

RESUMO Um estudo prospectivo, descritivo e longitudinal foi realizado de setembro de 2018 a Julho 2019 para determinar a microbiologia do abscesso anal como preditivo de fístulas anais em pacientes que compareceram à consulta externa da unidade de Coloproctologia do Hospital Central da Universidade Dr. Antonio María Pineda. Neste estudo, a população foi composta por pacientes com diagnóstico de abscesso anal, sem comorbidades ou contraindicações associadas à cirurgia, que concordaram em participar do estudo. Uma amostra intencional não probabilística, composta por 42 pacientes foi determinada. Um protocolo de estudo com base na consulta em regime ambulatorial foi aplicado aos pacientes que atenderam os critérios de inclusão estabelecidos, para realizar a anamnese, o exame físico e a devida cultura da supuração do abscesso anal para posteriormente estabelecer o comportamento médico e cirúrgico. Os resultados foram expressos em números absolutos e porcentagens, observando-se a prevalência de abscessos isquiorretais, seguidos por abscessos profundos no espaço pós-anal. Bactérias anaeróbias foram isoladas em 100% das amostras. Em todos os pacientes com fístulas, E. Coli foi isolada como um germe predominante.

Application of 16S rRNA Gene-Targeted Next-Generation Sequencing for Bacterial Pathogen Detection in Continuous Ambulatory Peritoneal Dialysis Peritonitis / 대한임상미생물학회지

BACKGROUND: 16S rRNA gene-targeted next-generation sequencing (NGS) can detect microorganisms in a comprehensive reference database. To date, NGS has been successfully applied to samples such as urine, blood, and synovial fluid. However, there is no data for continuous ambulatory peritoneal dialysis (CAPD) fluid. The purpose of this study was to evaluate the clinical usefulness of microbiome analysis of CAPD fluids for the diagnosis of CAPD peritonitis.METHODS: We included 21 patients with high suspicion of CAPD peritonitis. Routine CAPD fluid culture was performed using a pellet of 50 mL CAPD fluid onto the chocolate and blood agar for two days, and thioglycollate broth for one week. 16S rRNA gene-targeted NGS of pellets, stored at −70℃ was performed with MiSeq (Illumina, USA).RESULTS: Many colonized or pathogenic bacteria were detected from CAPD fluids using NGS and the microbiomes were composed of 1 to 29 genera with a cut-off 1.0. Compared to the culture results, NGS detected the same pathogens in 6 of 18 valid results (three samples failed with low read count). Additionally, using NGS, anaerobes such as Bacteroides spp. and Prevotella spp. were detected in six patients. In two of five samples in which no bacterial growth was detected, possible pathogens were detected by NGS.CONCLUSION: To our knowledge, this is the first report about the application of 16S rRNA gene-targeted NGS for diagnosis of CAPD peritonitis. Etiology of culture-negative CAPD peritonitis can be better defined in NGS. Furthermore, it also helped the detection of anaerobic bacteria.

Contamination of paper points used by students during preclinical and clinical endodontic procedures / Contaminação de cones de papel utilizados por estudantes durante procedimentos endodônticos pré-clínicos e clínicos

Objective: To determine the presence of facultative anaerobic bacteria in the paper points used by students and to perform a pilot test to determine whether sterilization of these materials influences their absorption capacity. Material and Methods: This study consists of two phases. The first is a descriptive phase where a representative sample of paper points (n = 72) was collected from the students and information about the points was voluntarily contributed. The points were placed in saline solution after they were collected, and mechanically shaken for 60 s. Then, 300 IU of the solution was seeded on blood agar in duplicates and incubated for 5 days under anaerobic conditions. The second phase was experimental, during which five paper points of each of the existing sizes were sterilized (Numbers: 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 70, and 80), and their capacity to absorb water was compared with that of the control or non-sterilized points. Results: The study determined that 22% (n=16) of the points were primarily contaminated by gram-positive bacilli, followed by gram-positive cocci, among which Staphylococcus epidermidis was identified. The presence of contamination was not significantly associated with the conditions of the paper points (p > 0.05). Furthermore, no significant effect (p>0.05) on the absorption capacity of these materials was detected in the sterilization test. Conclusion: Contamination was observed in the paper points used by the students, confirming the importance of implementing sterilization protocols. The sterilization protocols implemented in this study did not affect the absorption capacity of the points. (AU)

Objetivo: Determinar a presença de bactérias anaeróbias facultativas nos cones de papel usados pelos alunos e realizar um teste piloto para determinar se a esterilização desses materiais influencia sua capacidade de absorção. Material e Métodos: Este estudo consiste em duas fases. A primeira é uma fase descritiva, em que uma amostra representativa de cones de papel (n = 72) foi coletada dos alunos e informações sobre os cones foram voluntariamente fornecidas. Os cones foram colocados em solução salina após serem coletados e agitados mecanicamente por 60 s. Em seguida, 300 µl da solução foram semeadas em ágar em duplicatas e incubadas por 5 dias em condições anaeróbicas. A segunda fase foi experimental, durante a qual cinco pontos de papel de cada um dos tamanhos existentes (Números: 15, 20, 25, 30, 35, 40,45, 50, 55, 60, 70 e 80) foram esterilizados e suas respectivas capacidades de absorção de água foram comparadas com a do controle ou dos cones de papel não esterilizados. Resultados: O estudo determinou que 22% (n = 16) dos pontos estavam primariamente contaminados por bacilos gram positivos, seguidos por cocos gram-positivos, dentre os quais o Staphylococcus epidermidis foi identificado. A presença de contaminação não foi significativamente associada às condições dos cones de papel (p> 0,05). Além disso, nenhum efeito significativo (p> 0,05) sobre a capacidade de absorção desses materiais foi detectado no teste de esterilização. Conclusão: A contaminação foi observada nos cones de papel utilizados pelos estudantes,confirmando a importância da implementação de protocolos de esterilização.Os protocolos de esterilização implementados neste estudo não afetaram a capacidade de absorção dos cones (AU)

Comparison of Total Anaerobic Microbiota in Periodontitis Before and After the Subgingival Irrigation with Chlorhexidine at 0. 12 % / Comparación de la Microbiota Anaerobia Total en Periodontitis antes y Después de la Irrigación Subgingival con Clorhexidina al 0, 12 %

ABSTRACT: The objective of this study was to determine the effect of the subgingival irrigation of chlorhexidine 0.12 % of the total anaerobic microbiota. Microbial sampling to 30 subjects with periodontitis stage II Grade B, in pockets with a periodontal probing depth > 4 mm. The subgingival irrigation was made with 5 mL of chlorhexidine in the test group and with 5 mL of distilled water in the control group. 24 hours after the procedure was obtained a second sample to compare. It was found that the subgingival irrigation with chlorhexidine at 0.12 % achieved a statistically significant decrease in anaerobic microbiota (p< 0.05).

RESUMEN: El objetivo del presente estudio fue determinar el efecto de la irrigación subgingival de la clorhexidina 0,12 % sobre la microbiota anaeróbica total. Se tomaron muestras microbiológicas a 30 sujetos con periodontitis estadio II grado B, en sacos periodontales con una profundidad de sondaje > 4 mm. Se realizó la irrigación subgingival con 5 mL. de clorhexidina en el grupo test y con 5 mL. de agua destilada en el grupo control. 24 horas después del procedimiento se obtuvo una segunda muestra a comparar. Se detectó que la irrigación subgingival con clorhexidina al 0,12 % logra disminuir en forma estadísticamente significativa la microbiota anaeróbica total (p< 0,05).

Recuperación de bacterias aerobias y anaerobias de pacientes con apendicitis aguda mediante botellas de hemocultivo / Recovery of aerobic and anaerobic bacteria from patients with acute appendicitis using blood culture bottles

Introducción. La apendicitis aguda es la primera causa de abdomen agudo; sin embargo, poco se conoce sobre las bacterias asociadas y su perfil de sensibilidad. Objetivo. Identificar y determinar el patrón de resistencia de las bacterias aerobias y anaerobias aisladas en cultivo de líquido periapendicular tomado de los pacientes con apendicitis aguda, y establecer la proporción de bacterias según la fase clínica. Materiales y métodos. Se llevó a cabo un estudio descriptivo y prospectivo en el Hospital Universitario de San José de Bogotá (Colombia), en pacientes mayores de 16 años sometidos a apendicectomía abierta. Se tomaron muestras de líquido periapendicular, las cuales se sembraron directamente en botellas de hemocultivos para aerobios y anaerobios. Resultados. Se incluyeron 154 pacientes. Del total de cultivos, el 87 % (n=134) fueron positivos: 77 % (n=118) para aerobios y 51 % (n=79) para anaerobios. La proporción de cultivos positivos fue inferior en los casos de apendicitis no complicada, en comparación con aquellos de apendicitis complicada (80 % (66/83) Vs. 95 % (67/71); p=0,003). Los microorganismos aislados con mayor frecuencia fueron: Escherichia coli (53 %) (n=84), Bacteroides sp. (25 %) (n=25), Propionibacterium acnes (21 %) (n=21), Staphylococci coagulasa negativo (17 %) (n=27), Enterococcus sp. (10 %) (n=15) y Fusobacterium sp. (11 %) (n=11). La sensibilidad de E. coli a la amplicilina sulbactam fue de 30 %. La sensibilidad de Bacteroides spp. a la clindamicina y la ampicilina sulbactam fue de 91 %. El 100 % de los anaerobios fueron sensibles a piperacilina tazobactam, ertapenem, meropenem y metronidazol. Conclusiones. Los cultivos intraoperatorios son pertinentes en la apendicitis para determinar el patrón epidemiológico local, y establecer los antibióticos profilácticos y terapéuticos para esta enfermedad. Su siembra directa en botellas de hemocultivo permite una gran recuperación de microorganismos.

Introduction: Acute appendicitis is the first cause of acute abdomen, however, there is a little information about the associated bacteria and its sensibility profile. Objetive: To identify and to determine the resistance pattern of aerobic and anaerobic bacteria isolated in periapendicular fluid cultures taken in patients with acute appendicitis and to establish the proportions of isolates according to the clinical phase. Materials and methods: A descriptive and prospective study was undertaken at the Hospital Universitario de San José (Bogotá, Colombia) of patients older than sixteen years of age, undergoing an open appendectomy. A sample of periappendiceal fluid was taken, which was deposited directly into aerobic and anaerobic blood culture bottles. Results: One hundred and fifty-four patients were included. The overall positivity of cultures was 87% (n=1344); 77% (n=118) for aerobes and 51% (n=79) for anaerobes. The proportion of positive cultures was lower in the uncomplicated appendicitis cases as compared to the complicated ones (80% (66/83) vs. 95%(67/71), p = 0.003). The microorganisms isolated most frequently were: Escherichia coli (53%) (n=84); Bacteroides spp. (25%) (n=25); Propionibacterium acnes (21%) (n=21); coagulase negative Staphylococci (17%) (n=27); Enterococcus spp. (11%) (n=15), and Fusobacterium spp. (11%) (n=11). The sensitivity of E.coli to ampicillin/sulbactam was 30%. The sensitivity of Bacteroides spp. to clindamycin and ampicillin/sulbactam was 91%. All anaerobe isolates were sensitive to piperacillin/tazobactam, ertapenem, meropenem and metronidazole. Conclusions: Intraoperative cultures in acute appendicits are relevant in order to determine the local epidemiological pattern and to establish prophylactic and therapeutic antibiotics for this pathology; direct inoculation in blood culture bottles allows a high recovery of microorganisms.

Subgingival bacterial microbiota associated with ovine periodontitis / Microbiota bacteriana subgengival associada à periodontite ovina

Periodontitis is an inflammatory response in a susceptible host caused by complex microbiota, predominantly composed of Gram-negative anaerobic bacteria. Aiming to characterize the subgingival bacterial microbiota associated with ovine periodontitis, polymerase chain reaction (PCR) was performed in subgingival periodontal pocket samples of 14 sheep with severe periodontitis and in subgingival sulcus biofilm of 14 periodontally healthy sheep in search mainly of Gram-negative and Gram-positive microorganisms considered important periodontopathogens. The most prevalent bacteria in the sheep with periodontal lesions were Tannerella forsythia (78.6%), Treponema denticola (78.6%), Fusobacterium nucleatum (64.3%), and Porphyromonas gingivalis (50%), whereas in the healthy sheep, F. nucleatum (42.8%) was the most often detected bacterium. Statistically significant differences were observed for Campylobacter rectus, Enterococcus faecium, Prevotella nigrescens, T. forsythia, and T. denticola (p<0.05) in the sheep with periodontitis in the comparison between groups. Aggregatibacter actinomycetemcomitans, Enterococcus faecalis, and Porphyromonas gulae were not detected in any of the samples analyzed. In conclusion, C. rectus, E. faecium, P. nigrescens, T. forsythia, and T. denticola were associated with severe lesions caused by ovine periodontitis, and F. nucleatum was the most prevalent microorganism in the subgengival sulcus biofilm of healthy sheep.(AU)

Periodontite é a resposta inflamatória de um hospedeiro suscetível causada por complexa microbiota, composta predominantemente por bactérias anaeróbias Gram-negativas. Com o objetivo de caracterizar a microbiota bacteriana subgengival associada à periodontite ovina foi realizada a reação em cadeia da polimerase (PCR) de amostras de biofilme subgengival de 14 ovinos com a enfermidade e 14 ovinos periodontalmente saudáveis, com destaque para micro-organismos Gram-negativos e Gram-positivos considerados importantes periodontopatógenos. As bactérias mais prevalentes em 14 animais com lesões periodontais foram Tannerella forsythia (78,6%), Treponema denticola (78,6%), Fusobacterium nucleatum (64,3%) e Porphyromonas gingivalis (50%). Entretanto, nos 14 ovinos sem lesões periodontais, F. nucleatum (42,8%) foi a bactéria mais detectada. Associação estatisticamente diferente foi observada para Campylobacter rectus, Enterococcus faecium, Prevotella nigrescens, T. forsythia e T. denticola (p<0,05) nos ovinos com periodontite em comparação entre os dois grupos. Aggregatibacter actinomycetemcomitans, Enterococcus faecalis e Porphyromonas gulae não foram detectados em nenhuma das amostras pesquisadas. Conclui-se que C. rectus, E. faecium, P. nigrescens, T. forsythia e T. denticola estão associados às lesões resultantes da periodontite ovina com manifestação clínica grave e F. nucleatum o micro-organismo mais prevalente no biofilme subgengival de animais periodontalmente sadios.(AU)

Insights into the microbial community structure of anaerobic digestion of municipal solid waste landfill leachate for methane production by adaptive thermophilic granular sludge

BACKGROUND: The amount of municipal solid waste (MSW) gradually increased along with the rapid development of modern cities. A large amount of landfill leachate are generated with excessive chemical oxygen demand (COD), which create a great deal of pressure on the environment-friendly treatment process. Anaerobic digestion is an ideal technique to solve the above problem. RESULTS: A thermophilic granular sludge was successfully adapted for anaerobic digestion of MSW leachate (from an aging large-scale landfill) for methane production. The COD degradation efficiency improved by 81.8%, while the methane production rate reached 117.3 mL CH4/(g VS d), which was 2.34-fold more than the control condition. The bacterial and archaeal communities involved in the process were revealed by 16S rRNA gene high-throughput pyrosequencing. The richness of the bacterial community decreased in the process of thermophilic granular sludge, while the archaeal community structure presented a reverse phenomenon. The bacterial genus, Methanosaeta was the most abundant during the mesophilic process, while Methanobacterium, Methanoculleus, Methanosaeta and Methanosarcina were more evenly distributed. The more balanced community distribution between hydrogenotrophic and acetotrophic methanogens implied a closer interaction between the microbes, which further contributed to higher methane productivity. The detailed relationship between the key functional communities and anaerobic digestion performances were demonstrated via the multivariate canonical correspondence analysis. Conclusions: With the assistance of adaptive thermophilic granular sludge, microbial community structure was more evenly distributed, while both of COD degradation rate and methane production was improved during anaerobic digestion of MSW landfill leachate.

Clinical evaluation of cetylpyridinium chloride-containing mouthwash in halitosis

Halitosis is a very common disease that affects the majority of the population and is characterized by unpleasant odor during expiration. Anaerobic bacteria produce a range of malodorous substances including volatile sulfur compounds. To reduce oral malodor, the amount of oral microorganisms should be managed through brushing, scraping, and use of antibacterial agents. In this study, a mouthwash containing 0.05% cetylpyridinium chloride was tested on 22 candidates with oral malodor for two weeks to confirm oral malodor reduction through the use of antibacterial mouthwashes. Volatile sulfur compound measurements were significantly lower after using the mouthwash than before using it; thus, the mouthwash effectively reduced oral malodor.

Concomitant Liver and Brain Abscesses Caused by Parvimonas Micra / 대한소화기학회지

Anaerobic infections have been reported to be responsible for 3–10% of pyogenic liver abscesses in Korea, and reported anaerobes include Fusobacterium, Bacillus fragilis, and Bacteroides melaninogenicus. Parvimonas micra is an anaerobic, Gram-positive, non-spore-forming bacterial species and a constituent of normal flora on skin, vagina, gastrointestinal tract, and oral cavity that can cause opportunistic infections. However, it has only rarely been reported to be a cause of liver abscess; only one such case has been reported in Korea. We experienced a case of concomitant liver and brain abscesses caused by Parvimonas micra in a non-immunodeficient 65-year-old female patient without diabetes or periodontal disease. Parvimonas micra infection was confirmed by blood culture using VITEK® 2 cards and by bacterial 16s ribosomal RNA gene sequencing. We conclude that we should not overlook anaerobes as a cause of liver abscess.

Antibiotic sensitivity pattern of bacterial isolates in patients with chronic rhinosinusitis in Kaduna, Nigeria

Introduction: A common practice in the management of patients with chronic rhinosinusitis (CRS), the empirical use of antibiotics may contribute to treatment failure and to the development of antimicrobial resistance. Objective: To determine the antibiotic sensitivity pattern of aerobic and anaerobic bacteria associated with CRS. Methods: This was a prospective cross-sectional study in which endoscopically guided middlemeatal swabs (IBMSpss, version 16.0, Chicago, IL, USA) were aseptically taken from patients diagnosed with CRS after obtaining informed consent and ethical clearance. The samples were sent to the laboratory for qualitative and semiquantitative analysis via gram stain, aerobic, anaerobic cultures and antibacterial sensitivity tests. The collected data was analyzed using SPSS for Windows, version 16 (SPSS Inc, Chicago, IL, USA). Simple statistical parameters and paired sample t-test were used, as appropriate. Results: There were 74 (56.92%) bacterial growths, out of which 55 (74.32%) were aerobic and 19 (25.68%) were anaerobic isolates, from a total of 130 patients. About 13 (17.5%- 18%) of these bacterial growths yielded a mixed growth of aerobic and anaerobic isolates. Themost common bacterial isolates were 26 (35.14%) Staphylococcus aureus, Haemophilus influenzae 9 (12.16%), Streptococcus viridians 8 (10.81%), and Streptococcus pneumoniae 5 (6.76%). Augmentin, ciprofloxacin, and Peflacine were found to bemost effective, followed by levofloxacin, Rocephin, erythromycin and Zinat in that order (AU)

Occurrence of salmonella spp. in broiler chicken feces in the central region of the state of Minas Gerais, Brazil

Bacteria of the genus Salmonella are gram-negative rods, facultative anaerobic non-spore formers, which may or may not be flagellated. These bacteria are frequently detected in poultry products and are important in animal and public health. The purpose of this study was to investigate the occurrence of Salmonella spp. in feces from broiler chicken litters located in municipalities of the central region of the state of Minas Gerais, Brazil. The study analyzed 845 feces samples from different poultry farms collected between September 2016 and March 2017. Pre-enrichment broths, specific selective broths and selective indicator agar were used for the analyses. The suspected isolates were submitted to biochemical testing and serotyping with "O" and "H" antisera to identify the Salmonella serotypes. The results showed that 213 feces samples were contaminated with Salmonella, and the serotypes found were: S. Minnesota, S. Sandiego, S. Schwarzengrund, S. Infantis, S. Hadar, S. enterica subsp. enterica (O: 4.5), S. Montevideo, S. Miami, S. Heidelberg, S. Cerro, S. Ndolo, S. Panama, S. Anatum, S. Tennessee, S. Agona, S. Newport and S. Muenster. The Minnesota serotype was predominant among the isolates investigated. Actions to improve the biosafety of commercial poultry farms are necessary to prevent possible contamination of poultry products that endanger human health.

As bactérias do gênero Salmonella são bastonetes gram-negativos, formadores anaeróbicos facultativos não esporos, que podem ou não ser flagelados. Essas bactérias são freqüentemente detectadas em produtos avícolas e são importantes na saúde animal e pública. O objetivo deste estudo foi investigar a ocorrência de Salmonella spp. em fezes de ninhadas de frangos de corte localizados em municípios da região central do estado de Minas Gerais, Brasil. O estudo analisou 845 amostras de fezes de diferentes granjas de aves coletadas entre setembro de 2016 e março de 2017. Caldos de pré-enriquecimento, caldos seletivos específicos e ágar indicador seletivo foram utilizados para as análises. Os isolados suspeitos foram submetidos a testes bioquímicos e sorotipagem com anti-soros "O" e "H" para identificar os sorotipos de Salmonella. Os resultados mostraram que 213 amostras de fezes estavam contaminadas com Salmonella, e os sorotipos encontrados foram: S. Minnesota, S. Sandiego, S. Schwarzengrund, S. Infantis, S. Hadar, S. enterica subsp. enterica (O: 4.5), S. Montevidéu, S. Miami, S. Heidelberg, S. Cerro, S. Ndolo, S. Panamá, S. Anatum, S. Tennessee, S. Agona, S. Newport e S. Muenster . O sorotipo Minnesota foi predominante entre os isolados investigados. Ações para melhorar a biossegurança das granjas comerciais de aves são necessárias para evitar a possível contaminação dos produtos avícolas que põem em risco a saúde humana.

Epidemiología de bacterias anaerobias aisladas en muestras clínicas en el hospital san juan de dios, San José, Costa Rica, durante el trienio 2014, 2015 y 2016 / Epidemiology of anaerobic bacteria isolated in clinical samples at San Juan de Dios Hospital, San José, Costa Rica, during the 2014, 2015 and 2016 triennium

Resumen La epidemiología de las posibles poblaciones en riesgo de sufrir una infección por bacterias anaerobias a nivel nacional es desconocida, se debería de incentivar el conocimiento en los servicios de salud sobre este tipo de infecciones. Las bacterias anaerobias están relacionadas en los medios hospitalarios como causa importante de morbilidad, razón por la cual es conveniente conocer la epidemiologia y prevalecía de especies involucradas. En el Laboratorio de Bacteriología del Hospital San Juan de Dios, durante tres años, se analizaron un total de 1545 pacientes sospechosos de microorganismos anaerobios en medios prerreducidos, mediante un tamizaje se redujeron a un total de 469 posibles muestras, las cuales, fueron enviadas al Laboratorio de Investigación en Bacteriología Anaerobia (LIBA) para su correspondiente identificación. A lo largo de las semanas epidemiológicas de los tres años se encontraron en promedio de 1.77 casos confirmados / semana, con razón de sexo positiva a favor de los masculinos. Se determinó que solo 245 de las muestras enviadas presentaban uno o varios microorganismos anaerobios estrictos representando un 15.85% del total, identificándose 39 especies diferentes, en 306 cepas aisladas. Las mayormente importante fue el género Bacteriodes, provenientes de cavidad abdominal seguido de abscesos y heridas de piel. El presente estudio tiene como objetivo presentar datos que respalden la importancia clínica de la búsqueda de microorganismos anaerobios y que ayuden a los analistas de bacteriología a guiar cuales son los principales microorganismos esperables en muestras clínicas, además de conocer la prevalencia en general.

Abstract The epidemiology of the possible populations at risk of suffering an infection by anaerobic bacteria a national level is unknown, it should be encouraged the knowledge in the health services about this type of infections. Anaerobic bacteria are related in hospital environments as an important cause of morbidity, which is why it is convenient to know the epidemiology and prevalence of species involved. In the Bacteriology Laboratory of the Hospital San Juan de Dios, for three years, a total of 1545 patients suspected of anaerobic microorganisms in prereduced media were analyzed, through a screening was reduced to a total of 469 possible samples, which were sent to the Anaerobic Bacteriology Research Laboratory (LIBA) for its corresponding identification. Throughout the epidemiological weeks of the three years were found on average of 1.77 confirmed cases / week, with a positive sex ratio in favor of men. It was determined that only 245 of the samples sent had one or several strict anaerobic microorganisms representing 15.85% of the total, identifying 39 different species, in 306 isolated strains. The most important was the genus Bacteriodes, coming from the abdominal cavity followed by abscesses and skin wounds. The present study aims to present data that support the clinical importance of the search for anaerobic microorganisms and that help the analysts of bacteriology to guide which are the main expected microorganisms in clinical samples, in addition to knowing the prevalence in general.

Revisión de los efectos del consumo de alcohol sobre la salud periodontal / A review of the effects of alcohol consumption on periodontal health

La periodontitis es una enfermedad infecciosa caracterizada por la formación de bolsas periodontales que alojan microorganismos patógenos y por la inflamación de los tejidos de soporte dentario, ambas condiciones suman sus efectos nocivos provocando resorción ósea alveolar y deterioro del resto de los tejidos periodontales. La enfermedad se inicia por acumulación de placa bacteriana, pero su patogénesis progresa asociada a la respuesta inmune/inflamatoria del huésped que incrementa el deterioro de los tejidos periodontales. El alcohol es la sustancia de abuso de mayor consumo en todo el mundo. La literatura presenta numerosos estudios de correlación entre consumo de alcohol y desarrollo de enfermedades orales en seres humanos, aunque algunos autores han reportado efectos benéficos del consumo moderado de determinados tipos de alcohol. En este trabajo, luego de una revisión exhaustiva de la literatura, se concluye que el consumo abusivo de alcohol daña los tejidos periodontales y aumenta la predisposición de desarrollar periodontitis. Para finalizar, se describen los principales mecanismos que podrían estar implicados (AU)

Periodontitis is an infectious disease characterized by the formation of periodontal pockets that harbor pathogenic microorganisms and by the inflammation of dental support tissues, both of which add their harmful effects causing alveolar bone resorption and deterioration of the rest of the periodontal tissues. The disease was initiated by the accumulation of bacterial plaque, but its pathogenesis progresses associated with the immune / inflammatory response of the host that increases the deterioration of the periodontal tissues. Alcohol is the most commonly abused substance in the world. The literature presents numerous studies of the correlation between alcohol consumption and development of oral diseases in humans, although some authors have reported beneficial effects of moderate consumption of certain types of alcohol. In this paper, after an exhaustive review of the literature, it is concluded that the abusive consumption of alcohol damages the periodontal tissues and increases the predisposition to develop periodontitis. Finally, the main mechanisms that could be involved were described (AU)

Microbial load after selective and complete caries removal in permanent molars: a randomized clinical trial

Abstract The aim of this randomized clinical trial was to compare the remaining microbial load after treatments based on complete and selective caries removal and sealing. Patients with active carious lesions in a permanent molar were randomly allocated into 2 groups: a test group (selective caries removal-SCR; n=18) and a control group (complete caries removal - CCR; n=18). Dentin samples were collected following the excavation and three months after sealing. Streptococcus species, Streptococcus mutans, Lactobacillus species, and total viable microorganisms were cultured to count the viable cells and frequency of species isolation. CCR resulted in significant lower total viable microorganisms counts (p≤0.001), Streptococcus species (p≤0.001) and Lactobacillus species (p≤0.001) initially. However, after sealing, a decrease in total viable microorganisms, Streptococcus species, and Lactobacillus species in the SCR resulted in no difference between the groups after 3 months. In conclusion, selective caries removal is as effective as complete caries removal in reducing dentin bacterial load 3 months after sealing.

Resumo O objetivo deste ensaio clínico randomizado foi comparar os microrganismos remanescentes após tratamentos baseados em remoção total de tecido cariado e selamento e a remoção seletiva de tecido cariado e selamento. Pacientes com lesões de cárie ativas em molares permanentes foram divididos aleatoriamente em dois grupos: grupo teste (remoção seletiva de tecido cariado-SCR; n=18), e grupo de controle (remoção total de tecido cariado-CCR; n=18). Amostras de dentina foram obtidas após a remoção da tecido cariado e após 3 meses de selamento das cavidades. Streptococcus spp., Streptococcus mutans, Lactobacillus spp. e microrganismos viáveis totais foram cultivados para contagem de células e frequência de isolamento de espécies. CCR resultou em menores contagens totais de microorganismos viáveis (p≤0,001), Streptococcus spp. (p≤0,001) e Lactobacillus spp. (p≤0,001) inicialmente. Entretanto, após o selamento, uma redução significativa nas contagens totais de microrganismos viáveis, Streptococcus spp. e Lactobacillus spp. resultou em nenhuma diferença entre os grupos após 3 meses. Conclui-se que a remoção seletiva de cárie é tão seletiva quanto a remoção completa de cárie na redução da infecção dentinária após três meses com selamento da lesão.

Changes in Antimicrobial Susceptibility Pattern of Blood Isolates at a University Hospital in the Kyungnam area during 2005–2014 / 고신대학교의과대학학술지

OBJECTIVES: Blood culture is a one of the most important procedure for diagnosis and treatment of infectious disease, but distribution of pathogenic species and the antimicrobial susceptibility can be vary from pathogen, individual trait, regional or environmental features. In this study, we investigated the changes in frequency of occurrence and antimicrobial susceptibility pattern of blood isolates from 2005 to 2014. METHODS: Data of blood isolates from Kosin Gospel Hospital during 2005 to 2014 were analyzed retrospectively. Blood isolates were cultured for 5 days using BACTEC Plus Aerobic/F and BACTEC lytic/10 Anaerobic/F. Identification and antimicrobial susceptibility test was performed using VITEK 1 system, VITEK 2 XL, PHOENIX 100 and conventional method. RESULTS: 9,847 isolates were identified during 10 years. Among the isolates aerobic or falcutative anaerobic bacteria were isolated in 99.5% specimens, anaerobic were 0.1%, and fugi were 0.4%. Most commonly isolated bacteria were coagulase-negative Staphylococcus (CoNS) followed by Escherichia coli, Staphylococcus aureus and Klebsiella pneumoniae. Candida parapsilosis were most frequently isolated among fungi. The proportion of S. aureus, A. baumannii and E. faecium were increased, while Pseudomonas aeruginosa and Streptococcus pneumoniae decreased over decennium. Imipenem resistant K. pneumoniae were identified. Vancomycin resistant E. faecium and imipenem resistant A. baumannii were increased (7.1% in 2005 to 12.3% in 2014, 0% in 2005 to 55.6% in 2014, respectively). CONCLUSIONS: Over the last 10 year, CoNS were the most frequently isolated pathogen. Imipenem resistant K. pneumoniae was emerged. Vancomycin resistant E. faecium and imipenem resistant A. baumannii increased during this period.

Potencial antimicrobiano de extratos glicólicos vegetais sobre bactérias anaeróbias / Antimicrobial potential of plant glycolic extracts on anaerobic bacteria

A resistência microbiana aos antibióticos disponíveis é preocupação constante, devido à dificuldade no tratamento de infecções causadas por cepas resistentes, em decorrência do uso indiscriminado de antimicrobianos. Assim, a busca por terapias antimicrobianas alternativas tem sido crescente e necessária, sendo a fitoterapia umas das opções de escolha. O objetivo do presente estudo foi analisar a atividade antibacteriana de extratos glicólicos de Achyrocline satureioides (macela), Cynara scolymus (alcachofra), Hamamelis virginiana (hamamelis) e Persea americana (abacateiro), pelos períodos de 5 min e 24 h de exposição sobre bactérias anaeróbias Fusobacterium nucleatum subsp. nucleatum, Parvimonas micra, Porphyromonas endodontalis e Porphyromonas gingivalis, em culturas planctônica e biofilmes. As bactérias armazenadas a -80ºC foram ativadas em caldo Brucella enriquecido (hemina 1%, menadiona 1% e sangue de carneiro desfibrinado 5%) e incubadas em câmara de anaerobiose por 48 h a 37ºC por sete dias. A partir de culturas puras, o teste de microdiluição em caldo foi conduzido em microplacas por meio de suspensões bacterianas padronizadas em solução fisiológica estéril (NaCl 0,9%) e diluições dos extratos em caldo, sendo as placas incubadas por 48 h a 37ºC em anaerobiose. Alíquotas de cada poço foram semeadas em ágar Brucella enriquecido. Após incubação, a Concentração Inibitória Mínima (CIM) e Concentração Bactericida Mínima (CBM) foram determinadas. As concentrações efetivas de cada extrato foram aplicadas sobre os biofilmes de cada espécie, formados em microplacas a partir de suspensões bacterianas puras padronizadas na escala 0,5 de McFarland. As microplacas foram incubadas por sete dias a 37ºC para formação dos biofilmes, sendo o meio trocado a cada 48 h. Os biofilmes foram tratados por 5 min e 24 h. Em seguida, foram lavados e desprendidos por homogeneizador ultrassônico. As suspensões diluídas foram adicionadas em ágar Brucella enriquecido. Após 48 h, as Unidades Formadoras de Colônia por mililitro (UFC/mL) foram determinadas. Os resultados foram analisados por ANOVA e teste de Tukey ou por Kruskal-Wallis e teste Dunns, ambos com nível de significância de 5% (p≤0,05). Sobre as culturas planctônicas, a CIM e CBM dos extratos foi determinada apenas para F. nucleatum. A CBM dos extratos de A. satureioides, C. scolymus e P. americana foi obtida sobre P. micra. Não foi obtida atividade bactericida para P. endodontalis e P. gingivalis. Sobre biofilmes, todas as espécies apresentaram reduções significativas quando expostas aos extratos em ambos os tempos. Pode-se concluir que os extratos testados apresentaram efeito bacteriostático sobre F. nucleatum. Atividade bactericida dos extratos foi observada sobre F. nucleatum, bem como sobre P. micra, exceto para H. virginiana. Os extratos avaliados também apresentaram efeito antibiofilme sobre F. nucleatum, P. micra, P. endodontalis e P. gingivalis por 5 min e 24 h de exposição(AU)

Microbial resistance to antibiotics available is constant concern, due to the difficulty in treating infections caused by resistant strains as a result of the indiscriminate use of antimicrobials. Thus, the search for antimicrobial alternative therapies has been growing and necessary, being one option the herbal medicine. The objective of the present study was to analyze the antibacterial activity to Achyrocline satureioides glycolic extracts (macela), Cynara scolymus (artichoke), Hamamelis virginiana (Witch-Hazel) and Persea americana (avocado), for periods of 5 min and 24 h from exhibition on anaerobic bacteria Fusobacterium nucleatum subsp. nucleatum, Parvimonas micra, Porphyromonas gingivalis and Porphyromonas endodontalis in planktonic communities and biofilms. Bacteria stored at -80°C have been activated in Brucella broth enriched (hemin 1%, menadione 1% and defibrinated sheep blood 5%) and incubated in anaerobiose chamber for 48 h at 37° C for seven days. From pure cultures, the microdiluição test in broth was conducted in microplates through standardized bacterial suspensions in sterile saline solution (NaCl 0.9%) and dilution of the extracts in broth, being incubated plates for 48 h at 37° C in anaerobiosis. Aliquots of each well were sown in Brucella agar enriched. After incubation, the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were determined. Effective concentrations of each extract were applied on the biofilms of each species, formed in microplates from pure bacterial suspensions in 0.5 McFarland scale standard. The microplates were incubated for 7 days at 37°C for the formation of biofilms, being the culture medium replaced every 48 h. Biofilms were treated for 5 min and 24 h have been washed and given off by ultrasonic homogenizer. Dilute suspensions were added in Brucella agar enriched. After 48 h, the Colony Forming Units per milliliter (CFU/ml) were determined. The results were analyzed by ANOVA and Tukey test, or Kruskal-Wallis test and Dunns, both with a significance level of 5% (p ≤ 0.05). On the planktonic cultures, CIM and CBM of extracts was determined only to F. nucleatum. The CBM of the extracts of A. satureioides, C. scolymus and P. americana was obtained on P. micra. Bactericidal activity was not obtained for P. endodontalis and P. gingivalis. About biofilms, all species exhibited significant reductions when exposed to the extracts in both times. It can be concluded that the extracts tested showed bacteriostatic effect on F. nucleatum. Bactericidal activity of extracts was observed on F. nucleatum and P. micra, except for H. virginiana. The extracts evaluated also presented antibiofilme effect on F. nucleatum, P. micra, P. endodontalis and P. gingivalis for 5 min and 24 h(AU)