Molecular characterization of Aeromonas hydrophila isolates from diseased fishes in district Kasur, Punjab, Pakistan / Caracterização molecular de isolados de Aeromonas hydrophila de peixes doentes no distrito de Kasur, Punjab, Paquistão

Abstract Pakistan is an agricultural country and fisheries play a very important role in the economic development of the country. Different diseases are prevalent in Pakistani fish but information related to the causative agents is not well-known. Keeping in view the significance of bacterial pathogens as the causative agents of multiple fish diseases, the present study was conducted for identification, characterization and analysis of virulence genes of Aeromonas spp. isolated from diseased fishes. A total of fifty fish samples having multiple clinical indications were collected from different fish farms of district Kasur, Punjab Pakistan. For isolation of Aeromonas spp. samples were enriched and inoculated on Aeromonas isolation medium. Isolates were identified and characterized by different biochemical tests, Analytical Profile Index (API) 20E kit and Polymerase Chain Reaction (PCR) assays. All isolates were screened for three putative virulence genes including aerolysin (aer), haemolysin (hyl) and heat labile cytotonic enterotoxin (alt). Seven isolates of Aeromonas (A.) hydrophila were retrieved and identified based on API 20E. These isolates were further confirmed as A. hydrophila on the basis of PCR assays. Three isolates were detected positive for the presence of virulence genes (alt and hyl). Whereas aerolysin (aer) gene was not present in any of A. hydrophila isolates. The present study confirmed A. hydrophila as the causative agent of epizootic ulcerative syndrome and motile Aeromonas septicemia in fish farms of district Kasur, Punjab Pakistan. Moreover, detection of two virulence genes (alt and hyl) in A. hydrophila isolates is a threat for fish consumers of study area.

Resumo O Paquistão é um país agrícola, onde a pesca desempenha um papel muito importante para o desenvolvimento econômico. Diferentes doenças são prevalentes em peixes do Paquistão, mas as informações relacionadas aos agentes causadores não são bem conhecidas. Tendo em vista a importância dos patógenos bacterianos como agentes causadores de múltiplas doenças em peixes, o presente estudo foi conduzido para identificação, caracterização e análise de genes de virulência de isolados de Aeromonas spp. de peixes doentes. Foram coletadas 50 amostras de peixes com múltiplas indicações clínicas em diferentes fazendas do distrito de Kasur, Punjab, Paquistão. Para isolar Aeromonas spp., as amostras foram enriquecidas e inoculadas em meio de isolamento. Os isolados foram identificados e caracterizados por diferentes testes bioquímicos, kit Analytical Profile Index (API) 20E, e ensaios de reação em cadeia da polimerase (PCR). Todos os isolados foram selecionados para três genes de virulência putativos, incluindo aerolisina (aer), hemolisina (hyl) e enterotoxina citotônica termolábil (alt). Sete isolados de Aeromonas hydrophila foram recuperados e identificados com base no API 20E. Esses isolados foram posteriormente confirmados como A. hydrophila de acordo com ensaios de PCR. Três isolados indicaram a presença de genes de virulência (alt e hyl), enquanto o gene aerolisina (aer) não esteve presente em nenhum dos isolados de A. hydrophila. O presente estudo confirmou A. hydrophila como o agente causador da síndrome ulcerativa epizoótica e septicemia móvel por Aeromonas em fazendas de peixes, no distrito de Kasur, Punjab, Paquistão. Além disso, a detecção de dois genes de virulência (alt e hyl) em isolados de A. hydrophila é uma ameaça para os consumidores de peixes da área de estudo.

Efficacy and safefy of Polymyxin B treatment for neutropenic patients suffering from refractory Gram-negative bacterial bloodstream infection / 中华血液学杂志

Objective: To assess the efficacy and safety of polymyxin B in neutropenic patients with hematologic disorders who had refractory gram-negative bacterial bloodstream infection. Methods: From August 2021 to July 2022, we retrospectively analyzed neutropenic patients with refractory gram-negative bacterial bloodstream infection who were treated with polymyxin B in the Department of Hematology of the First Affiliated Hospital of the Soochow University between August 2021 to July 2022. The cumulative response rate was then computed. Results: The study included 27 neutropenic patients with refractory gram-negative bacterial bloodstream infections. Polymyxin B therapy was effective in 22 of 27 patients. The median time between the onset of fever and the delivery of polymyxin B was 3 days [interquartile range (IQR) : 2-5]. The median duration of polymyxin B treatment was 7 days (IQR: 5-11). Polymyxin B therapy had a median antipyretic time of 37 h (IQR: 32-70). The incidence of acute renal dysfunction was 14.8% (four out of 27 cases), all classified as "injury" according to RIFLE criteria. The incidence of hyperpigmentation was 59.3%. Conclusion: Polymyxin B is a viable treatment option for granulocytopenia patients with refractory gram-negative bacterial bloodstream infections.

Evaluation of three alum-precipitated Aeromonas hydrophila vaccines administered to Labeo rohita, Cirrhinus mrigala and Ctenopharyngodon idella: immunokinetics, immersion challenge and histopathology / Avaliação de três vacinas de Aeromonas hydrophila precipitadas com alúmen administradas a Labeo rohita, Cirrhinus mrigala e Ctenopharyngodon idella: imunocinética, desafio de imersão e histopatologia

Aeromonas hydrophila is a cause of infectious disease outbreaks in carp species cultured in South Asian countries including Pakistan. This bacterium has gained resistance to a wide range of antibiotics and robust preventive measures are necessary to control its spread. No prior use of fish vaccines has been reported in Pakistan. The present study aims to develop and evaluate inactivated vaccines against local strain of A. hydrophila in Pakistan with alum-precipitate as adjuvant. The immunogenic potential of vaccine was evaluated in two Indian major carps (Rohu: Labeo rohita, Mori: Cirrhinus mrigala) and a Chinese carp (Grass carp: Ctenopharyngodon idella). Fish were vaccinated intraperitoneally followed by a challenge through immersion. Fish with an average age of 4-5 months were randomly distributed in three vaccinated groups with three vaccine concentrations of 108, 109 and 1010 colony forming unit (CFU)/ml and a control group. Fixed dose of 0.1ml was applied to each fish on 1st day and a booster dose at 15 days post-vaccination (DPV). Blood samples were collected on 14, 28, 35, 48 and 60 DPV to determine antibody titers in blood serum using compliment fixation test (CFT). Fish were challenged at 60 DPV with infectious A. hydrophila with 108 CFU/ml through immersion. Significantly higher levels of antibody titers were observed from 28 DPV in all vaccinated groups as compared to those in the control group. In challenge experiment the average RPS (relative percent survivability) was 71% for groups vaccinated with 109 and 1010 CFU/ml and 86% for 108 CFU/ml. Vaccine with 108 CFU/ml induced highest immune response followed by 109 and 1010 CFU/ml. The immune response of L. rohita and C. idella was better than that of C. mrigala. In general, normal histopathology was [...].

Aeromonas hydrophila é uma causa de surtos de doenças infecciosas em espécies de carpas cultivadas em países do sul da Ásia, incluindo o Paquistão. Essa bactéria ganhou resistência a uma ampla gama de antibióticos, e medidas preventivas robustas são necessárias para controlar sua disseminação. Nenhum uso anterior de vacinas para peixes foi relatado no Paquistão. O presente estudo tem como objetivo desenvolver e avaliar vacinas inativadas contra cepa local de A. hydrophila no Paquistão com precipitado de alúmen como adjuvante. O potencial imunogênico da vacina foi avaliado em duas carpas principais indianas (Rohu: Labeo rohita, Mori: Cirrhinus mrigala) e uma carpa chinesa (Grass Carp: Ctenopharyngodon idella). Os peixes foram vacinados por via intraperitoneal, seguido de um desafio por imersão. Peixes com idade média de 4-5 meses foram distribuídos aleatoriamente em três grupos vacinados com três concentrações de vacina de 108, 109 e 1010 unidades formadoras de colônias (UFC) / ml e um grupo de controle. Foi aplicada dose fixa de 0,1ml em cada peixe no 1º dia e dose de reforço 15 dias pós-vacinação (DPV). Amostras de sangue foram coletadas em 14, 28, 35, 48 e 60 DPV para determinar os títulos de anticorpos no soro sanguíneo usando o teste de fixação de elogio (CFT). Os peixes foram desafiados a 60 DPV com infecciosa A. hydrophila com 108 CFU / ml por imersão. Níveis significativamente mais elevados de títulos de anticorpos foram observados em 28 DPV em todos os grupos vacinados, em comparação com aqueles no grupo de controle. Na experiência de desafio, o RPS médio (sobrevivência percentual relativa) foi de 71% para os grupos vacinados com 109 e 1010 CFU / ml e 86% para 108 CFU / ml. A vacina com 108 UFC / ml induziu a maior resposta imune seguida por 109 e 1010 UFC / ml. A resposta imune de L. rohita e C. idella foi melhor do que a de C. mrigala. Em geral, histopatologia normal foi observada em diferentes [...].

Temporal association between antibiotic use and resistance in Gram-negative bacteria / Associação temporal entre o uso de antibióticos e resistência em bactérias gram-negativas

Abstract The β-lactam/lactamase inhibitors (BLBLIs) combination drugs are considered an effective alternative to carbapenems. However, there is a growing concern that the increased use of BLBLIs may lead to increased resistance. This study determined the temporal association between the consumption of BLBLI and the antimicrobial resistance in Gram-negative bacteria. In this retrospective study, electronic data on the Gram-negative bacterial isolates, including A. baumannii, P. aeruginosa, E. coli, and K. pneumoniae from in-patients and susceptibility testing results were retrieved from the medical records of the clinical laboratory. A linear regression and cross-correlation analysis were performed on the acquired data. Increasing trends (p<0.05) in the consumption of BIBLI and carbapenem with a median use of 27.68 and 34.46 DDD/1000 PD per quarter were observed, respectively. A decreased trend (p=0.023) in the consumption of fluoroquinolones with a median use of 29.13 DDD/1000 PD per quarter was observed. The resistance rate of K. pneumoniae was synchronized with the BIBLI and carbapenem consumptions with a correlation coefficient of 0.893 (p=0.012) and 0.951 (p=0.016), respectively. The cross-correlation analysis against the consumption of BIBLI and meropenem resistant K. pneumoniae was peaked at 0-quarter lag (r=951, p=0.016). There was an increasing trend in the consumption of BLBLI and carbapenems. The increasing trend in the rates of resistance to piperacillin/tazobactam, in line with the increasing consumption of BLBLI, suggests that BLBLI has to be used with caution and cannot be directly considered as a long-term alternative to carbapenems.

Resumo Os medicamentos combinados de β-lactâmicos / inibidores da lactamase (BLBLIs) são considerados uma alternativa eficaz aos carbapenêmicos. No entanto, existe uma preocupação crescente de que o aumento do uso de BLBLIs pode levar ao aumento da resistência. Este estudo determinou a associação temporal entre o consumo de BLBLI e a resistência antimicrobiana em bactérias gram-negativas. Neste estudo retrospectivo, os dados eletrônicos sobre as bactérias gram-negativas isoladas, incluindo A. baumannii, P. aeruginosa, E. coli e K. pneumoniae de pacientes internados e os resultados dos testes de suscetibilidade foram recuperados dos registros médicos do laboratório clínico. Uma regressão linear e análise de correlação cruzada foram realizadas nos dados adquiridos. Foram observadas tendências crescentes (p < 0,05) no consumo de BIBLI e carbapenem com uma mediana de uso de 27,68 e 34,46 DDD/1000 PD por trimestre, respectivamente. Foi observada uma tendência de diminuição (p = 0,023) no consumo de fluoroquinolonas com uma mediana de uso de 29,13 DDD/1000 PD por trimestre. A taxa de resistência de K. pneumoniae foi sincronizada com os consumos de BIBLI e carbapenem com coeficiente de correlação de 0,893 (p = 0,012) e 0,951 (p = 0,016), respectivamente. A análise de correlação cruzada contra o consumo de BIBLI e K. pneumoniae resistente ao meropenem atingiu o pico no intervalo de 0 quarto (r = 951, p = 0,016). Houve uma tendência de aumento no consumo de BLBLI e carbapenêmicos. A tendência crescente nas taxas de resistência a piperacilina/tazobactam, em linha com o consumo crescente de BLBLI, sugere que BLBLI deve ser usado com cautela e não pode ser considerado diretamente como alternativa de longo prazo aos carbapenêmicos.

Evaluation of three alum-precipitated Aeromonas hydrophila vaccines administered to Labeo rohita, Cirrhinus mrigala and Ctenopharyngodon idella: immunokinetics, immersion challenge and histopathology / Avaliação de três vacinas de Aeromonas hydrophila precipitadas com alúmen administradas a Labeo rohita, Cirrhinus mrigala e Ctenopharyngodon idella: imunocinética, desafio de imersão e histopatologia

Abstract Aeromonas hydrophila is a cause of infectious disease outbreaks in carp species cultured in South Asian countries including Pakistan. This bacterium has gained resistance to a wide range of antibiotics and robust preventive measures are necessary to control its spread. No prior use of fish vaccines has been reported in Pakistan. The present study aims to develop and evaluate inactivated vaccines against local strain of A. hydrophila in Pakistan with alum-precipitate as adjuvant. The immunogenic potential of vaccine was evaluated in two Indian major carps (Rohu: Labeo rohita, Mori: Cirrhinus mrigala) and a Chinese carp (Grass carp: Ctenopharyngodon idella). Fish were vaccinated intraperitoneally followed by a challenge through immersion. Fish with an average age of 4-5 months were randomly distributed in three vaccinated groups with three vaccine concentrations of 108, 109 and 1010 colony forming unit (CFU)/ml and a control group. Fixed dose of 0.1ml was applied to each fish on 1st day and a booster dose at 15 days post-vaccination (DPV). Blood samples were collected on 14, 28, 35, 48 and 60 DPV to determine antibody titers in blood serum using compliment fixation test (CFT). Fish were challenged at 60 DPV with infectious A. hydrophila with 108 CFU/ml through immersion. Significantly higher levels of antibody titers were observed from 28 DPV in all vaccinated groups as compared to those in the control group. In challenge experiment the average RPS (relative percent survivability) was 71% for groups vaccinated with 109 and 1010 CFU/ml and 86% for 108 CFU/ml. Vaccine with 108 CFU/ml induced highest immune response followed by 109 and 1010 CFU/ml. The immune response of L. rohita and C. idella was better than that of C. mrigala. In general, normal histopathology was observed in different organs of vaccinated fish whereas minor deteriorative changes were found in fish vaccinated with higher concentrations of the vaccine.

Resumo Aeromonas hydrophila é uma causa de surtos de doenças infecciosas em espécies de carpas cultivadas em países do sul da Ásia, incluindo o Paquistão. Essa bactéria ganhou resistência a uma ampla gama de antibióticos, e medidas preventivas robustas são necessárias para controlar sua disseminação. Nenhum uso anterior de vacinas para peixes foi relatado no Paquistão. O presente estudo tem como objetivo desenvolver e avaliar vacinas inativadas contra cepa local de A. hydrophila no Paquistão com precipitado de alúmen como adjuvante. O potencial imunogênico da vacina foi avaliado em duas carpas principais indianas (Rohu: Labeo rohita, Mori: Cirrhinus mrigala) e uma carpa chinesa (Grass Carp: Ctenopharyngodon idella). Os peixes foram vacinados por via intraperitoneal, seguido de um desafio por imersão. Peixes com idade média de 4-5 meses foram distribuídos aleatoriamente em três grupos vacinados com três concentrações de vacina de 108, 109 e 1010 unidades formadoras de colônias (UFC) / ml e um grupo de controle. Foi aplicada dose fixa de 0,1ml em cada peixe no 1º dia e dose de reforço 15 dias pós-vacinação (DPV). Amostras de sangue foram coletadas em 14, 28, 35, 48 e 60 DPV para determinar os títulos de anticorpos no soro sanguíneo usando o teste de fixação de elogio (CFT). Os peixes foram desafiados a 60 DPV com infecciosa A. hydrophila com 108 CFU / ml por imersão. Níveis significativamente mais elevados de títulos de anticorpos foram observados em 28 DPV em todos os grupos vacinados, em comparação com aqueles no grupo de controle. Na experiência de desafio, o RPS médio (sobrevivência percentual relativa) foi de 71% para os grupos vacinados com 109 e 1010 CFU / ml e 86% para 108 CFU / ml. A vacina com 108 UFC / ml induziu a maior resposta imune seguida por 109 e 1010 UFC / ml. A resposta imune de L. rohita e C. idella foi melhor do que a de C. mrigala. Em geral, histopatologia normal foi observada em diferentes órgãos de peixes vacinados, enquanto pequenas alterações deteriorantes foram encontradas no grupo de controle e nos peixes vacinados com concentrações mais altas da vacina.

Mortalidad en niños con bacteriemia por Stenotrophomonas maltophilia: revisión sistemática / Mortality in children with Stenotrophomonas maltophilia-related bacteremia: A systematic review

Introducción: En los niños, la bacteriemia por Stenotrophomonas maltophilia es considerada una complicación severa y asociada a una elevada mortalidad. Con el objetivo de conocer la mortalidad asociada a esa condición, se realizó una revisión sistemática de la literatura. Material y métodos: Se aplicó una estrategia de búsqueda bibliográfica con las palabras clave: bacteriemia por Stenotrophomonas maltophilia, niños y adolescentes como únicos filtros. Se informan la mediana y los valores intercuartílicos de la frecuencia de la mortalidad reportada por los estudios incluidos. Resultados: Se identificaron 165 estudios potencialmente útiles. De ellos, se seleccionaron finalmente, 9 estudios para ser incluidos. La incidencia de mortalidad a consecuencia de una bacteriemia por S.maltophilia fue del 25%; Q25: 11­Q75: 36; rango: 6,06 a 40,6. Consideraciones finales: La bacteriemia por Sm tuvo un alto porcentaje de mortalidad en especial en pacientes con patología subyacente y uso de procedimientos invasivos y el uso inadecuado de antibióticos empíricos (AU)

Introduction: In children, Stenotrophomonas maltophilia-related bacteremia is considered a severe complication associated with high mortality. With the aim to determine the mortality associated with this condition, a systematic review of the literature was conducted. Material and methods: A literature search strategy was applied using the keywords: bacteremia due to Stenotrophomonas maltophilia, children, and adolescents as the only filters. The median and interquartile ranges of the mortality rates described in the studies included are reported. Results: A total of 165 potentially useful studies were identified, of which nine were finally selected to be included in the analysis. The incidence of S.maltophilia bacteremia-related mortality was 25%; Q25: 11­Q75: 36; range: 6.06 to 40.6. Final considerations: S.maltophilia-related bacteremia was associated with a high mortality rate especially in patients with an underlying disease, when invasive procedures were performed, and when emperical antibiotics were inadequately used (AU)

Blue and red light photoemitters as approach to inhibit Staphylococcus aureus and Pseudomonas aeruginosa growth / Fotoemissores de luz azul e vermelha como abordagem para inibir o crescimento de Staphylococcus aureus e Pseudomonas aeruginosa

The ability of pathogenic bacteria acquire resistance to the existing antibiotics has long been considered a dangerous health risk threat. Currently, the use of visible light has been considered a new approach to treat bacterial infections as an alternative to antibiotics. Herein, we investigated the antimicrobial effect of two range of visible light, blue and red, on Staphylococcus aureus and Pseudomonas aeruginosa, two pathogenic bacterial commonly found in healthcare settings-acquired infections and responsible for high rate of morbidity and mortality. Bacterial cultures were exposed to blue or red light (470 nm and 660 nm) provided by light-emitting diodes - LED. The fluencies and irradiance used for blue and red light were 284.90 J/cm², 13.19 mW/cm² and 603.44 J/cm², 27.93 mW/cm² respectively. Different experimental approaches were used to determine the optimal conditions of light application. Only exposure to blue light for 6 hours was able to inhibit about 75% in vitro growth of both bacterial species after 24 hours. The surviving exposed bacteria formed colonies significantly smaller than controls, however, these bacteria were able to resume growth after 48 hours. Blue light was able to inhibit bacterial growth upon inoculation in both saline solution and BHI culture medium. We can conclude that blue light, but not red light, is capable of temporarily retarding the growth of gram negative and gram positive bacteria.

A capacidade das bactérias patogênicas adquirirem resistência aos antibióticos existentes há muito tempo é considerada uma ameaça perigosa à saúde. Atualmente, o uso da luz visível tem sido considerado uma nova abordagem no tratamento de infecções bacterianas como alternativa aos antibióticos. Neste trabalho, investigamos o efeito antimicrobiano de duas faixas de luz visível, azul e vermelha, em Staphylococcus aureus e Pseudomonas aeruginosa, duas bactérias patogênicas comumente encontradas em infecções adquiridas em instituições de saúde e responsáveis por alta taxa de morbimortalidade. As culturas bacterianas foram expostas à luz azul ou vermelha(470 nm e 660 nm) fornecida por diodos emissores de luz - LED. As fluências e irradiâncias utilizadas para luz azule vermelha foram 284,90 J/cm², 13,19 mW/cm² e 603,44 J/cm², 27,93 mW/cm², respectivamente. Várias abordagens experimentais foram utilizadas para determinar as condições ótimas de aplicação da luz. Apenas a exposição à luz azul por 6 horas foi capaz de inibir cerca de 75% o crescimento in vitro de ambas as espécies bacterianas após24 horas. As bactérias expostas sobreviventes formaram colônias com um tamanho significativamente menor do que os controles, contudo, essas bactérias conseguiram retomar o crescimento normal após 48 horas. A luz azul foi capaz de inibir o crescimento das bactérias após sua inoculação em solução salina ou no meio de cultura rico em nutrientes BHI. Podemos concluir que a luz azul mas não a luz vermelha é capaz de retardar temporariamente o crescimento de bactérias Gram-negativas e Gram-positivas.

Pseudotumor toraco-abdominal asociado a Bartonella henselae y Eikenella corrodens en un paciente pediátrico inmunocompetente / Thoraco-abdominal pseudotumor associated with Bartonella henselae and Eikenella corrodens in an immunocompetent patient

Resumen Los pseudotumores inflamatorios son poco frecuentes y escasamente descritos en la literatura y han sido asociados a infecciones polimicrobianas. Se presenta el caso de un niño de 9 años, procedente de Bolivia, quien consultó por dolor abdominal, baja de peso y vómitos, diagnosticándose un pseudotumor tóraco-abdominal. El laboratorio clínico, mediante el estudio de biología molecular en tejido, permitió la identificación de uno de los probables agentes etiológicos.

Abstract Inflammatory pseudotumors are a rare pathology and scarcely reported in the literature and have been associated with polymicrobial infections. Here, we present the case of a 9 years old boy from Bolivia, who presented with abdominal pain, weight loss and vomiting, who was diagnosed with a thoraco-abdominal pseudotumor. The micro-biology and molecular laboratories in tissue allowed the identification of one of the probable etiological agents.

Restricción programada de cefalosporinas de tercera generación en contexto de un brote de bacilos gramnegativos productores de beta-lactamasas tipo AmpC en unidades críticas: una experiencia de la vida real / Third-generation cephalosporins programmed restriction in the context of an outbreak of AmpC beta-lactamase-producing gram-negative bacilli in critical units: a real-life experience

INTRODUCCIÓN: La restricción programada (RP) de antimicrobianos puede disminuir selectivamente la tasa de infecciones por determinados microorganismos. En este sentido, los bacilos gramnegativos productores de beta-lactamasas AmpC (BGN-blaAmpC) son seleccionados por el sobreuso de cefalosporinas de tercera generación (C3G). Estas bacterias, también adquieren genes y co-producen otras beta-lactamasas, como las de Nueva Delhi (BGN-blaNDM). OBJETIVOS: Disminuir la tasa de aislamiento de BGN-blaAmpC y BGN-blaNDM en cultivos de pacientes de la UCI luego de una RP de C3G en el marco de un brote nosocomial por estos microrganismos. MATERIALES Y MÉTODOS: Estudio cuasi-experimental, previo (P1= 12 meses) y posterior (P2= 12 meses) a una RP de C3G en un hospital de adultos, donde, en el contexto de brote mencionado, se aplicaron medidas de control de infecciones generales. El uso de antimicrobianos se expresó como "porcentaje de los días de tratamiento (%DDT)"/100 camas ocupadas al día (100-COD). Se compararon las tasas de aislamiento de BGN-blaAmpC y BGN-blaNDM en hemocultivos (HC), mini-lavados bronquio-alveolares (mB) y urocultivos (UC) en la UCI. RESULTADOS: En P2 el consumo de C3G fue 2,5% DDT/100-COD. Hubo un descenso en los aislamientos de BGN-blaAmpC en HC (RR 0,48 [0,2-0,9] p < 0,02) y mB (RR 0,52 [0,3-0,9] p < 0,02), así como también de BGN-blaNDM en HC (RR 8,1 [1,6-39,4] p < 0,00). Conclusiones: La RP de C3G se asoció con la reducción de los BGN-blaAmpC y BGN-blaNDM en HC, así como de los BGN-blaAmpC mB.

BACKGROUND: Programmed restriction (PR) of antimicrobials can selectively decrease the rate of infections by certain microorganisms. In this sense, AmpC beta-lactamase-producing gram-negative bacilli (GNB-blaAmpC) are selected for the overuse of third generation cephalosporins (3GC). These bacteria also acquire genes and co-produce other β-lactamases, such as New Delhi ones (GNB-blaNDM). AIM: To decrease the isolation rate of GNB- blaAmpC and GNB- blaNDM in cultures from ICU patients after a PR of 3GC. METHODS: Quasi-experimental study, before (P1= 12 months) and after (P2= 12 months) a PR of 3GC in an adults' hospital. The use of antibiotics was expressed as "percentage days of treatment (%DOT)" /100 beds occupied per day (100-BOD). The rates of GNB-blaAmpC and GNB-blaNDM were compared in blood cultures (BC), mini-bronchio alveolar lavages (mB) and urine cultures (UC) in the ICU. RESULTS: In P2, 3GC consumption was 2.5% DOT/100-COD. There was a decrease in GNB-blaAmpC from BC (RR 0.48 [0.2-0.9] p < 0.02) and mB (RR 0.52 [0.3-0.9] p < 0.02), as well as of GNB-blaNDM from BC (RR 8.1 [1.6-39.4] p < 0.00). Conclusions: PR of 3GC was linked to the reduction of GNB-blaAmpC and GNB-blaNDM in BC, as well as GNB-blaAmpC in mB from ICU patients.

Prevalência de bastonetes Gram-negativos não fermentadores isolados em amostras de hemoculturas / Prevalence of non-fermenter Gram-negative bacilli isolated in blood culture samples

Objetivos: Avaliar a prevalência de bactérias não fermentadoras em amostras de hemoculturas provenientes das Unidades de Terapia Intensiva (UTI's) adulto e neonatal e da Unidade Coronariana (UC); definir o perfil de suscetibilidade aos antimicrobianos das cepas bacterianas prevalentes. Métodos: Foram coletados dados de todas as hemoculturas positivas das UTIs Adulto, Neonatal e UC de um hospital privado, em Juiz de Fora, Minas Gerais, Brasil, de janeiro de 2017 a janeiro de 2019. Resultados: Foram analisados 3.535 resultados de amostras de hemoculturas onde 2.464 (69,7%) foram negativas e 1.071 (30,3%) positivas para algum microrganismo. Dentre as amostras positivas foram encontrados 77 bastonetes Gram-negativos não fermentadores (6,9%), com a prevalência de Acinetobacter baumannii (51,9%) seguido de Pseudomonas aeruginosa (32,5%). As cepas de A. baumannii foram resistentes aos carbapenêmicos e às quinolonas. Quanto às cepas de P. aeruginosa, as drogas testadas que apresentaram maior resistência foram a ampicilina, ampicilina com tazobactam, as cefalosporinas de segunda e terceira geração, exceto ceftazidima; e a tigeciclina. As drogas que apresentaram boa atividade na inibição do crescimento das cepas analisadas foram tigeciclina para A. baumannii e colistina para ambas as cepas. Conclusão: o presente estudo alerta para a resistência a múltiplas classes de antimicrobianos das cepas advindas das UTIs e UC, demonstrando um cenário preocupante e a necessidade de desenvolvimento de novas drogas e novas medidas de controle.

Objectives: To evaluate the prevalence of non-fermenting bacteria in blood culture samples from the adult and neonatal intensive care units (ICUs) and the Coronary Care Unit (UC); define the antimicrobial susceptibility profile of prevalent bacterial strains. Methods: Data were collected on all positive blood cultures from the Adult, Neonatal and UC ICUs of a private hospital in Juiz de Fora, Minas Gerais, Brazil, from January 2017 to January 2019. Results: 3535 results of blood culture samples were analyzed, where 2464 (69.7%) were negative and 1071 (30.3%) positive for some microorganism. Among the positive samples, 77 non-fermenting Gram negative rods (6.9%) were found, with the prevalence of Acinetobacter baumannii (51.9%) followed by Pseudomonas aeruginosa (32.5%). A. baumannii strains were resistant to carbapenems and quinolones. As for strains of P. aeruginosa, the drugs tested that showed greater resistance were ampicillin, ampicillin with tazobactam, second and third generation of cephalosporins, except ceftazidime; and tigecycline. The drugs that showed good activity in inhibiting the growth of the strains analyzed were tigecycline for A. baumannii and colistin for both strains. Conclusion: the present study warns of resistance to multiple classes of antimicrobial strains from the ICUs and UC, demonstrating a worrying scenario and the need to develop new drugs and new control measures.

12 pasos para reducir la incidencia de carbapenemasas / 12 steps to mitigate the incidence of carbapenemases

La resistencia a los antimicrobianos es un grave problema para la salud mundial. Es aún más crítico en los hospitales debido a la aparición de bacterias Gram negativas resistentes a múltiples fármacos, asociadas a una alta mortalidad. Las opciones de tratamiento en estos casos son escasas, en general de alto costo. La alta densidad de consumo de antibióticos y la transmisión cruzada en este entorno amplifican este problema.Hay más evidencia del impacto de las medidas de control de infecciones que de las intervenciones de comités de antimicrobianos para mitigarlo. Además, pocos países cuentan con programas sólidos de control de infecciones para enfrentar este problema. En la presente revisión se propone una serie de 12 pasos a adoptar para mitigar la prevalencia de resistencia antimicrobiana y reducir la incidencia de carbapenemasas en las instituciones de salud. Estas recomendaciones deben interpretase como un ̈bundle ̈o paquete de medidas, en el cual todas son importantes. Aquellas que involucran la prevención de infecciones y/o colonizaciones y su diseminación son las de mayor impacto demostrado hasta ahora. Es esencial que los programas de optimización de uso de antimicrobianos cuenten con el empoderamiento de la conducción de las instituciones donde se lleven a cabo, así como también que estén constituidos por un equipo multidisciplinario eficiente, sólidamente entrenado, con metas y métricas objetivas y auditorias periódicas. También es recomendable que se incluyan recomendaciones para los tratamientos en pacientes en cuidados de fin de vida.

Antimicrobial resistance is a serious global health problem. It is even more critical in hospitals due to the emergence of multi-drug resistant Gram negative bacteria, associated with high mortality. The treatment options in these cases are scarce, generally high cost. The high density of antibiotic consumption and cross-transmission in this environment amplifies this problem.There is more evidence of the impact of Infection Control measures than of Antimicrobial Committee interventions to mitigate it. Furthermore, few countries have solid Infection Control programs to deal with this problem.This review proposes a series of 12 steps to adopt to mitigate the prevalence of antimicrobial resistance and reduce the incidence of carbapenemases in health institutions. These recommendations should be interpreted as a ̈Bundle ̈ or package of measures, in which all are important. Those that involve the prevention of infections and / or colonizations and their dissemination are the ones with the greatest impact demonstrated so far. It is essential that antimicrobial use optimization programs have the empowerment of the leadership of the institutions where they are carried out, as well as that they are constituted by an efficient multidisciplinary team, solidly trained, with objective goals and metrics and periodic audits. It is also recommended that recommendations be included for treatments in patients in end-of-life care.

Antimicrobial use for treatment of healthcare-associated infections and bacterial resistance in a reference neonatal unit

Abstract Objective The use of broad-spectrum antimicrobials, such as third and fourth-generation, are responsible for emergence of multidrug-resistant microorganisms in neonatal units. Furthermore, antimicrobial daily doses are not standardized in neonatology. This study aimed to investigate the association between the use of antimicrobial broad spectrum to bacterial sensitivity profile in a referral unit of neonatal progressive care. Methods This is a cohort study conducted in a referral neonatal progressive care unit from January 2008 to December 2016. The data of all hospitalized neonates was collected daily. The infection criteria used were the standardized national criteria, based on definitions of Center for Diseases Control and Prevention. In this study, the use of antimicrobials was evaluated as antimicrobial-day (ATM-day) and the ratio of multidrug-resistant microorganisms per 1000 ATM-day of broad spectrum was also calculated. The study was approved by the Institutional Review Board of the Universidade Federal de Minas Gerais (ETIC 312/08 e CAAE 58973616.2.0000.5149). Results From 2008 to 2016, 2751 neonates were hospitalized, corresponding to 60,656 patient-days. The ratio of multidrug-resistant microorganisms per 1000 ATM-day of broad spectrum was 1,3 in the first period and 4,3 in the second period (p = 0,005). Conclusion It was observed that use of broad-spectrum antimicrobials, especially those with coverage for Gram-negative bacteria, was associated with an increase of multidrug-resistant bacteria.

Infección por Sphingobacterium multivorum en un niño gran quemado / Sphingobacterium multivorum infection in a child with an extensive burn

Resumen Sphingobacterium multivorum es un bacilo gramnegativo no fermentador, que infrecuentemente causa enfermedad en humanos. En la literatura médica existen escasos reportes de infecciones causadas por este microorganismo, en general, en hospederos con alguna comorbilidad. Presentamos el primer caso reportado, según nuestro conocimiento, de una infección de piel y tejidos blandos por Sphingobacterium multivorum en un paciente pediátrico gran quemado.

Abstract Sphingobacterium multivorum is a non-fermentative gram-negative bacillus that rarely causes human infections. In the medical literature, the few reported cases of infections resulting from S. multivorum usually occurred in patients with an associated comorbidity. We present the first case report, according our knowledge, of a skin and soft tissue infection by S. multivorum infection in a pediatric patient after a burn injury.

Presencia de bacterias en prótesis dentales durante el proceso de elaboración / Presence of bacteria in dental prostheses during the elaboration process

Existe una creciente preocupación sobre el tema de la infección cruzada en clínicas y laboratorios dentales. El laboratorio odontológico debe seguir normas de bioseguridad que garanticen a todo el equipo de salud la prevención de estas infecciones. Los técnicos que allí laboran corren el riesgo de exponer su cara a salpicaduras, así como a rocíos de sangre y saliva. Este estudio fue diseñado para saber si los laboratorios a los que recurrimos cumplen con estas normas de bioseguridad, y qué tan confiados podemos estar de la desinfección por parte de ellos, ya que las prótesis deberían estar desinfectadas correctamente antes de colocarlas en boca (AU)

There is growing concern about the issue of cross infection in dental clinics and laboratories. The dental laboratory must follow biosafety standards that guarantee the prevention of these infections to the entire health team. The technicians who work there run the risk of exposing their face to splashes and spray of blood and saliva. This study was designed to find out if the laboratories we use comply with these biosafety standards, and how confident we can be of their disinfection by them, since the prostheses should be properly disinfected before placing them in the mouth (AU)

Compromiso del sistema nervioso central por Eikenella corrodens: revisión de la literatura médica a propósito de un caso clínico / Commitment of the central nervous system by Eikenella corrodens: literature review about a clinical case

Resumen Se presenta el caso de un paciente de 22 años sin antecedentes mórbidos que desarrolló un absceso cerebral secundario a una endocarditis por Eikenella corrodens. El diagnóstico se estableció mediante la detección por reacción de polimerasa en cadena universal en el líquido de la colección cerebral. La ecocardiografía transesofágica confirmó vegetaciones en la válvula mitral. Requirió la colocación de una derivación ventricular externa por vaciamiento del absceso a ventrículos cerebrales e hidrocefalia secundaria. Recibió 80 días de tratamiento antibacteriano efectivo. Su evolución fue favorable, con resolución completa de la infección verificada con imágenes y ecocardiografía de control. El seguimiento a los siete meses por una disfunción de válvula de drenaje ventrículo-peritoneal no demostró infección.

Abstract We present the case of a 22-year-old patient with no morbid history who developed a brain abscess secondary to endocarditis due to Eikenella corrodens. The diagnosis was established by detecting the microorganism by universal polymerase chain reaction in fluid from the brain collection. Trans-esophageal echocardiogram study confirmed vegetations in the mitral valve. External ventricular shunt placement was required due to emptying of abscess to brain ventricles and secondary hydrocephalus. He received 80 days of effective antibiotic treatment and his evolution was favorable with complete resolution of his infection verified with images and echocardiogram. Follow-up at seven months later due to ventricule-peritoneal drainage valve dysfunction did not confirmed infection.

Correlation of B-type Natriuretic Peptide Level with Hemodynamic Parameters and Inflammatory Cytokines in Patients with Gram-negative Sepsis / 中国医学科学院学报

Objective To study the correlation of B-type natriuretic peptide(BNP)level with hemodynamic parameters and inflammatory cytokines in patients with Gram-negative sepsis,and further determine the main factors for the significant increase of BNP level. Methods The prospective study method was applied,and septic patients infected with Gram-negative bacteria from May 2017 to October 2019 were enrolled.The patients were divided into the BNP<2400 ng/L group and the BNP≥2400 ng/L group by taking the average value of BNP as the dividing point.The independent predictors of BNP≥2400 ng/L were analyzed by Logistic regression.Pearson correlation analysis was used to analyze the correlation between BNP and various indicators. Results A total of 106 patients with Gram-negative sepsis were included,among which 60 cases present with higher serum BNP levels than the average of(2398.45 ± 421.45)ng/L.Thus BNP≥2400 ng/L was considered as a significantly increased BNP level.Multiple logistic regression analysis showed that cardiac index(CI)[odds ratio (

Active surveillance of carbapenem-resistant Gram-negative healthcare-associated infections in a low-middle-income country city

ABSTRACT Background: Carbapenem-resistance in healthcare-associated infections (HCAIs) is of great concern, and it is urgent to improve surveillance. We aimed to describe and analyze HCAIs trends on Gram-negative antimicrobial susceptibility in a city from a developing country, following the implementation of an active surveillance program. Methods: This is an aggregated study describing data from 24 hospitals with intensive care units, including a trend analysis by Joinpoint regression between January 2012 and December 2017. Results: There were 23,578 pathogens in 39,832 HCAIs, from which 16,225 were Gram-negatives (68.8%). Carbapenem susceptibility was lowest in A. baumannii (15.4-25.9%), K. pneumoniae (51.0-55.9%), and P. aeruginosa (64.9-84.1%) and highest in E. coli (96.5-99.2%). Only K. pneumoniae showed a significant Joinpoint at 95% confidence interval: −10.71% (−18.02; −2.75) from 2012 to 2014, p = 0.02, and 6.54% (−2.00; 15.83) from 2015 to 2017, p = 0.12, which was most influenced by urinary tract infections: −9.98% (−16.02; −3.48) from 2012 to 2014, p = 0.01, and 9.66% (−1.75; 22.39) from 2015 to 2017, p = 0.09. Conclusion: Although we found a significant change toward an improvement in carbapenem susceptibility in K. pneumoniae, resistance is high for most pathogens. These data should encourage health institutions to improve their prevention and control strategies.

Determinación de la contaminación bacteriana en conos de gutapercha en una clínica odontológica docente / Determination of bacterial contamination in gutapercha cones in an university dental clinic

Objetivo: El objetivo de este estudio fue determinar la contaminación bacteriana de los conos de gutapercha de tipo beta (ß) en los tiempos 0, 24, 47 y 72 horas de las diferentes proveedurías de la Clínica Odontológica de la Universidad Científica del Sur (Lima, 2020). Materiales y métodos: Se obtuvo 16 conos de gutapercha tipo beta (ß) de empaques cerrados bajo medidas asépticas, los cuales fueron colocados en viales con 2 ml de caldo BHI y, posteriormente, fueron sembrados en agar BHI, así como en medios selectivos agar manitol salado y agar MacConkey. Pasadas las 24 horas de incubación a 37 °C, se realizó la lectura de las placas y el conteo de UFC. El mismo procedimiento se realizó para los tiempos 24, 48 y 72 horas, lo que dio un total de 64 conos de gutapercha tipo ß. Resultados: Se observó que el nivel de contaminación bacteriana fue el mismo tanto entre las distintas proveedurías como a las 0, 24, 48 y 72 horas. Solo se hallaron diferencias estadísticamente significativas (p = 0,044) entre los distintos tiempos de la proveeduría número 5. Finalmente, todas las muestras sometidas a la prueba de la coagulasa arrojaron resultados negativos. Conclusión: Los conos de gutapercha de tipo beta (ß) se contaminaron por igual producto de su almacenamiento y manipulación, independientemente de la proveeduría en la que permanecieron. (AU)

Objective: The purpose of this study was to determinate the bacterial contamination of Beta (ß) gutta-percha cones at 0, 24, 47 and 72 hours of the different supplies of the Universidad Científica del Sur, Lima 2020. Materials and Methods: 16 ß-type gutta-percha cones were obtained of sealed packages under aseptic measurements, they were placed in vials with 2ml BHI and subsequently planted in BHI agar plates as well as in selective medias as Salted mannitol agar and MacConkey agar after 24 hours of incubation at 37 ° the plates were read and count in CFU, the same procedure was performed for the other times evaluated 24, 48 and 72 hours, giving a total of 64 ß-type gutta-percha cones. Results: It was observed that the level of bacterial contamination was the same among the different supplies in all the establish times of in this study 0, 24, 48 and 72 hours. Therefore, there were no significant differences in the level of bacterial contamination between the supplies. On the other hand, only statistically significant differences (p = 0.044) were found between the different times of the supply number 5. Finally, all the samples submitted to the coagulase test had a negative result. Conclusion: The gutta-percha cones of type ß were contaminated equally regardless of the supply in which they were stored or manipulated. (AU)

Uso racional de antibióticos y tecnología FilmArray para identificación rápida de bacteriemias en unidad de cuidados intensivos pediátrica / Rational use of antibiotics and FilmArray technology for rapid identification of bacteremias in a pediatric intensive care unit

INTRODUCCIÓN: Las infecciones graves son la principal causa de ingreso a cuidados intensivos pediátricos. El panel FilmArray BCID permite identificar rápidamente a microorganismos causantes de bacteriemias. OBJETIVO: evaluar la eficacia de la identificación rápida de microorganismos asociado a un Programa de Uso Racional de Antibióticos (URA) en reducir los tiempos de terapias antibióticas, en un hospital pediátrico. PACIENTES Y MÉTODO: Estudio retrospectivo, que incluyó 100 pacientes, en su primer episo dio de bacteriemia, divididos en 2 grupos de 50 cada uno: Intervención (FilmArray BCID y programa URA) y Controles históricos pareados para la misma especie del microrganismo identificado (microbiología convencional). Las variables evaluadas fueron los tiempos de identificación microbiana, latencia de la terapia dirigida y de desescalar antibióticos. RESULTADOS: Los grupos fueron comparables en características demográficas, foco de infección y etiología de bacteriemia. El tiempo promedio de identificación de microorganismos fue de 23 h (IC 95% 12,4-26,7) en el grupo intervención, y 70,5 h (IC 95% 65,2-78,6) en el control (p < 0,05), mientras que la latencia de inicio de terapia dirigida fue de 27,9 h (IC 95% 22,3-32,8) y 71,9 h (IC 95% 63,2-77,8) respectivamente (p < 0,05). El tiempo de desescalar o suspender antibióticos fue de 6,4 h (IC 95% 2,76-9,49) y 22 h (IC 95% 6,74-35,6) en los grupos mencionados (p > 0,05). CONCLUSIÓN: El panel FilmArray BCID articulado a un programa URA, contribuye a la identificación de los microorganismos causantes de bacteriemias en menor tiempo que los métodos convencionales, siendo una herramienta que optimiza las terapias antibióti cas en niños críticamente enfermos.

INTRODUCTION: Severe infections are the leading cause of admission to pediatric intensive care. The FilmArray BCID panel quickly identifies microorganisms that cause bacteremia. OBJECTIVE: To evaluate if the rapid identification of the microorganisms that cause bacteremia, along with a Rational Use of Antibio tics (RUA) Program, allows optimizing the time of antibiotic therapy in a pediatric hospital. PATIENTS AND METHOD: Retrospective study which included 100 patients presenting their first episode of bacteremia, divided into 2 groups of 50 each. The first one was Intervention (FilmArray BCID and RUA program) and the second one was Historical Controls (conventional automated ID/AST). The variables evaluated were the time required for microbial identification, duration of appropriate therapy, and antibiotic de-escalation. RESULTS: The groups were comparable in terms of demographic characteristics, focus of infection, and etiology of bacteremia. The average time of microorganisms' identification of the control group was 70.5 hours (IC 95% 65.2-78.6) and 23.0 hours (IC 95% 12.4 -26.7) in the intervention one (p < 0.05). The average time of targeted therapy onset was shorter in the intervention group (27.9 h [IC 95% 22.3-32.8]) than that of the control one (71.9 h [IC 95% 63.2-77.8]) (p < 0.05). Finally, the time to de-escalate or discontinue antibiotics in the intervention group and the control one was 6.4 hours (IC 95% 2.76-9.49) hours and 22.0 hours (IC 95% 6.74-35.6 h) respectively (p > 0.05). CONCLUSION: The FilmArray panel along with the RUA Program allows the identification of the microorganisms causing bacteremia faster than conventional methods, which positions it as a tool that optimizes antibiotic therapy of critical patients.

Predicting bacteria causing acute bacterial rhinosinusitis by clinical features / Predição de bactérias causadoras de rinossinusite bacteriana aguda por características clínicas

Abstract Introduction: Clinicians rely on clinical presentations to select therapeutic agents for acute bacterial rhinosinusitis. Streptococcus pneumoniae and Haemophilus influenzae are common in acute bacterial rhinosinusitis. Drug resistant Streptococcus pneumoniae and Haemophilus influenzae require different antibiotics. Objective: This study aimed to evaluate the associations between clinical features of acute bacterial rhinosinusitis and pathogenic bacteria. Methods: Sixty-four patients with acute bacterial rhinosinusitis were enrolled. Clinical features including nasal obstruction, discolored discharge, facial pain, smell disturbance, fever and laboratory findings of patients with acute bacterial rhinosinusitis were collected. The bacterial cultures of endoscopic middle meatal swabs were used as a reference. Results: Serum C-reactive protein level elevation correlated with the bacterial species (p = 0.03), by which was increased in 80.0% of Haemophilus influenzae rhinosinusitis and 57.1% of Streptococcus pneumoniae rhinosinusitis. The elevated C-reactive protein was the significant predictor for Haemophilus influenzae rhinosinusitis with the Odds Ratio of 18.06 (95% CI 2.36-138.20). The sensitivity of serum C-reactive protein level elevation for diagnosing Haemophilus influenzae rhinosinusitis was 0.80 (95% CI 0.49-0.94). Conclusion: Elevation of serum C-reactive protein level was associated with and predicted acute bacterial rhinosinusitis caused by Haemophilus influenzae.

Resumo: Introdução: Os médicos se baseiam nas características clínicas para a escolha dos agentes terapêuticos para o tratamento da rinossinusite bacteriana aguda. Streptococcus pneumoniae e Haemophilus influenzae são agentes comuns na rinossinusite bacteriana aguda. Streptococcus pneumoniae e Haemophilus influenzae resistentes a antibióticos requerem medicamentos diferentes. Objetivo: Avaliar as associações entre as características clínicas da rinossinusite bacteriana aguda e bactérias patogênicas. Método: O estudo incluiu 64 pacientes com rinossinusite bacteriana aguda. Foram coletadas e registradas as características clínicas, inclusive obstrução nasal, secreção com cor alterada, dor facial, distúrbios do olfato, febre e achados laboratoriais de pacientes com rinossinusite bacteriana aguda. As culturas bacterianas obtidas por swab endoscópico do meato médio foram usadas como referência. Resultados: A elevação do nível sérico de proteína C-reativa estava correlacionada com a espécie bacteriana (p = 0,03); ela estava aumentada em 80,0% das rinossinusites por Haemophilus influenzae e em 57,1% das rinossinusites por Streptococcus pneumoniae. A proteína C-reativa elevada foi um significativo fator preditor de rinossinusite por Haemophilus influenzae, com razão de probabilidade de 18,06 (IC 95% 2,36-138,20). A sensibilidade da elevação dos níveis séricos de proteína C-reativa para o diagnóstico de rinossinusite por Haemophilus influenzae foi de 0,80 (IC 95% 0,49 ± 0,94). Conclusão: A elevação dos níveis séricos de proteína C-reativa é um preditor de rinossinusite bacteriana aguda causada por Haemophilus influenzae.