Emergence of genetically unrelated NDM-1-producing Acinetobacter pittii strains in Paraguay

The New Delhi metallo-β-lactamase (NDM-1) was initially identified in Escherichia coli and Klebsiella pneumoniae isolates in Sweden, from a patient previously hospitalized in India.1 To date, NDM producers in Latin America have been scarce, and associated with species of Enterobacteriaceae from Guatemala, Mexico, Colombia and Brazil, although in Honduras it was reported in Acinetobacter baumannii.2?6 Here, we report two genetically unrelated NDM-1-producing Acinetobacter pittii isolates identified in Paraguay. Since 1996, the Pan American Health Organization (PAHO) has supported a regional surveillance system, the Antimicrobial Resistance Surveillance Network in Latin America (ReLAVRA), that includes 794 laboratories from 20 Latin American countries, including their respective reference laboratories.7 This network provides reliable, timely and reproducible microbiological data in order to improve patient care. A regional protocol for the detection of carbapenemases has been harmonized and implemented through ReLAVRA. Briefly, metallo-β-lactamase (MBL) production is suspected in isolates that exhibit decreased susceptibility to carbapenems (CLSI criteria) and a positive synergy test result between a disc containing 10 μg of imipenem and a disc containing 750 μg of EDTA plus 1900 μg of sodium thioglycolate.8 During 2012, following the ReLAVRA algorithm, the National Health Laboratory of Paraguay confirmed an MBL phenotype in two Acinetobacter spp. isolates recovered from a single hospital. This phenotype had not previously been observed in Acinetobacter spp. from Paraguay.Fil: Pasteran, Fernando. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; ArgentinaFil: Martinez Mora, Mario. Laboratorio Central de Salud Pública. Servicio Antimicrobianos; ParaguayFil: Albornoz, Ezequiel Pablo. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Faccone, Diego Francisco. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Franco, Rossana. Laboratorio Central de Salud Pública. Servicio Antimicrobianos; ParaguayFil: Ortellado, Juana. Universidad Nacional de Asunción; ParaguayFil: Melgarejo, Nancy. Laboratorio Central de Salud Pública. Servicio Antimicrobianos; ParaguayFil: Gómez, Sonia Alejandra. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Riquelme, Irma. Universidad Nacional de Asunción; ParaguayFil: Matheu, Jorge. Pan American Health Organization. Antimicrobial Resistance and Infection Control Program; Estados UnidosFil: Ramon Pardo, Jorge Matheu Pilar. Pan American Health Organization. Antimicrobial Resistance and Infection Control Program; Estados UnidosFil: Corso, Alejandra. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Enforcing Behavioral Profiles through Software-Defined Networks in the Industrial Internet of Things

The fourth industrial revolution is being mainly driven by the integration of Internet of Things (IoT) technologies to support the development lifecycle of systems and products. Despite the well-known advantages for the industry, an increasingly pervasive industrial ecosystem could make such devices an attractive target for potential attackers. Recently, the Manufacturer Usage Description (MUD) standard enables manufacturers to specify the intended use of their devices, thereby restricting the attack surface of a certain system. In this direction, we propose a mechanism to manage securely the obtaining and enforcement of MUD policies through the use of a Software-Defined Network (SDN) architecture. We analyze the applicability and advantages of the use of MUD in industrial environments based on our proposed solution, and provide an exhaustive performance evaluation of the required processes

Defining the Behavior of IoT Devices through the MUD Standard : Review, Challenges, and Research Directions

With the strong development of the Internet of Things (IoT), the definition of IoT devices' intended behavior is key for an effective detection of potential cybersecurity attacks and threats in an increasingly connected environment. In 2019, the Manufacturer Usage Description (MUD) was standardized within the IETF as a data model and architecture for defining, obtaining and deploying MUD files, which describe the network behavioral profiles of IoT devices. While it has attracted a strong interest from academia, industry, and Standards Developing Organizations (SDOs), MUD is not yet widely deployed in real-world scenarios. In this work, we analyze the current research landscape around this standard, and describe some of the main challenges to be considered in the coming years to foster its adoption and deployment. Based on the literature analysis and our own experience in this area, we further describe potential research directions exploiting the MUD standard to encourage the development of secure IoT-enabled scenarios

Faecal carriage of ESBL producing and colistin resistant Escherichia coli in avian species over a 2-year period (2017-2019) in Zimbabwe

DATA AVAILABILITY STATEMENT : The data presented in this study are deposited in the NCBI BioProject Number PRJNA 799483 with accession numbers listed in the Supplementary Datasheet 1.INTRODUCTION : Extended spectrum beta-lactamase (ESBL) producing Escherichia coli have become widespread among food producing animals. These strains serve as a reservoir of antibiotic resistance genes (ARGs) and act as a possible source of infection to humans as transmission can occur by direct or indirect contact. METHODS : This study investigated the faecal carriage of ESBL producing and colistin resistant E. coli in poultry over a 2-year period (2017-2019) from Zimbabwe. A total of 21 ESBL positive isolates from poultry cloacal specimens were selected for whole genome sequencing from animal E. coli isolates biobanked at the National Microbiology Reference laboratory using phenotypic susceptibility testing results from the National Escherichia coli Surveillance Program to provide representation of different geographical regions and year of isolation. Cloacal swabs were collected from 3000 broiler live birds from farm 1 and fromfarm2, 40 backyard chickens and 10 duckswere sampled. Antimicrobial susceptibility and ESBL testing were performed as per Clinical Laboratory Standards Institute guidelines. Whole genome sequencing of ESBL producing isolates was used to determine sequence types (STs), ARGs, and phylogroups. RESULTS : Twenty-one of the included E. coli isolates were confirmed as ESBL producers. Three defined sequence type clonal complexes (CCs) were identified (ST10CC, ST155CC and ST23CC), with ST10CC associated with the most antibiotic resistant profile. The ESBL phenotype was linked to the presence of either cefotaximase-Munich-14 (CTX-M-14) or CTX-M-79. Plasmid mediated quinolone resistant determinants identified were qnrB19 and qnrS1 and one ST10CC isolate from farm 1 broiler chickens harbored a mobile colistin resistance gene (mcr-1). Phylogenetic groups most identified were B1, A and unknown. DISCUSSIONS : The avian ESBL producing E. coli belonged to a diverse group of strains. The detection of several ARGs highlights the importance of implementing enhanced control measures to limit the spread in animals, environment, and humans. This is the first report of mcr-1 in Zimbabwe, which further underscores the importance of the One Health approach to control the spread and development of AMR.The National Health Laboratory Service (NHLS) and the University of Pretoria, South Africa, a strategic partnership between National Microbiology Reference Laboratory and Quadram BioSciences Institute.https://www.frontiersin.org/journals/cellular-and-infection-microbiologyam2023Medical Microbiolog

Molecular epidemiology of extended-spectrum beta-lactamase-producing extra-intestinal pathogenic Escherichia coli strains over a 2-year period (2017-2019) from Zimbabwe

This study was designed to characterize extended-spectrum beta-lactamase (ESBL)–producing extra-intestinal pathogenic Escherichia coli (E.coli) (ExPEC) associated with urinary tract infections in nine different geographic regions of Zimbabwe over a 2-year period (2017–2019). A total of 48 ESBL-positive isolates from urine specimen were selected for whole-genome sequencing from 1246 Escherichia coli isolates biobanked at the National Microbiology Reference laboratory using phenotypic susceptibility testing results from the National Escherichia coli Surveillance Programme to provide representation of different geographical regions and year of isolation. The majority of ESBL E. coli isolates produced cefotaximase-Munich (CTX-M)-15, CTX-M-27, and CTX-M-14. In this study, sequence types (ST) 131 and ST410 were the most predominant antimicrobial-resistant clones and responsible for the increase in ESBL–producing E. coli strains since 2017. Novel ST131 complex strains were recorded during the period 2017 to 2018, thus showing the establishment and evolution of this antimicrobial-resistant ESBL clone in Zimbabwe posing an important public health threat. Incompatibility group F plasmids were predominant among ST131 and ST410 isolates with the following replicons recorded most frequently: F1:A2:B20 (9/19, 47%), F2:A1: B (5/19, 26%), and F1:A1:B49 (8/13, 62%). The results indicate the need for continuous tracking of different ESBL ExPEC clones on a global scale, while targeting specific STs (e.g. ST131 and ST410) through control programs will substantially decrease the spread of ESBLs among ExPEC

PR-LncRNA Signature Regulates Glioma Cell Activity Through Expression of SOX Factors

Long non-coding RNAs (LncRNAs) have emerged as a relevant class of genome regulators involved in a broad range of biological processes and with important roles in tumor initiation and malignant progression. We have previously identified a p53-regulated tumor suppressor signature of LncRNAs (PR-LncRNAs) in colorectal cancer. Our aim was to identify the expression and function of this signature in gliomas. We found that the expression of the four PR-LncRNAs tested was high in human low-grade glioma samples and diminished with increasing grade of disease, being the lowest in glioblastoma samples. Functional assays demonstrated that PR-LncRNA silencing increased glioma cell proliferation and oncosphere formation. Mechanistically, we found an inverse correlation between PR-LncRNA expression and SOX1, SOX2 and SOX9 stem cell factors in human glioma biopsies and in glioma cells in vitro. Moreover, knock-down of SOX activity abolished the effect of PR-LncRNA silencing in glioma cell activity. In conclusion, our results demonstrate that the expression and function of PR-LncRNAs are significantly altered in gliomagenesis and that their activity is mediated by SOX factors. These results may provide important insights into the mechanisms responsible for glioblastoma pathogenesis.PA, JA-I and AS-A were recipients of a predoctoral fellowship from the Spanish Association Against Cancer (AECC Gipuzkoa), Basque Government and Instituto Salud Carlos III. This work was supported by grants from the Carlos III Institute of Health and the European Regional Development Fund (PI13/02277, CP16/00039, DTS16/084, and PI16/01580) and Industry and Health Departments of the Basque Country

Emergence of NDM-1-producing Klebsiella pneumoniae in Guatemala

Fil: Pasteran, Fernando. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas; Argentina.Fil: Albornoz, Ezequiel. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas; Argentina.Fil: Faccone, Diego. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas; Argentina.Fil: Gómez, Sonia. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas; Argentina.Fil: Valenzuela, Claudia. Unidad Central de Referencia para la Vigilancia Epidemiológica. Laboratorio Nacional de Salud. Sección Bacteriología; Guatemala.Fil: Morales, Melissa. Unidad Central de Referencia para la Vigilancia Epidemiológica. Laboratorio Nacional de Salud. Sección Bacteriología; Guatemala.Fil: Estrada, Pavela. Hospital Infantil de Infectología y Rehabilitación; Guatemala.Fil: Valenzuela, Laura. Hospital General San Juan de Dios; Ciudad de Guatemala.Fil: Matheu, Jorge. Pan American Health Organization/World Health Organization. Alert and Response and Epidemic Diseases; Estados Unidos.Fil: Guerriero, Leonor. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas; Argentina.Fil: Arbizú, Enrique. Unidad Central de Referencia para la Vigilancia Epidemiológica. Laboratorio Nacional de Salud. Sección Bacteriología; Guatemala.Fil: Calderón, Yeraldine. Unidad Central de Referencia para la Vigilancia Epidemiológica. Laboratorio Nacional de Salud. Sección Bacteriología; Guatemala.Fil: Ramon-Pardo, Pilar. Pan American Health Organization/World Health Organization. Alert and Response and Epidemic Diseases; Estados Unidos.Fil: Corso, Alejandra. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas; Argentina

Capability of national reference laboratories in Latin America to detect emerging resistance mechanisms

Fil: Corso, Alejandra. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Bacteriología. Servicio Antimicrobianos; Argentina.Fil: Guerriero, Leonor. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Bacteriología. Servicio Antimicrobianos; Argentina.Fil: Pasterán, Fernando. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Bacteriología. Servicio Antimicrobianos; Argentina.Fil: Ceriana, Paola. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Bacteriología. Servicio Antimicrobianos; Argentina.Fil: Callejo, Raquel. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Bacteriología. Servicio Bacteriología Especial; Argentina.Fil: Prieto, Mónica. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Bacteriología. Servicio Bacteriología Especial; Argentina.Fil: Tuduri, Ezequiel. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Bacteriología. Servicio Antimicrobianos; Argentina.Fil: Lopardo, Horacio. Hospital de Pediatría S.A.M.I.C. "Prof Dr. Juan Garrahan"; Argentina.Fil: Vay, Carlos. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Hospital de Clínicas "José de San Martín"; Argentina.Fil: Smayevsky, Jorgelina. Centro de Educación Médica e Investigaciones Clínicas "Norberto Quirno"; Argentina.Fil: Tokumoto, Marta. Fundación Favaloro; Argentina.Fil: Alvarez, Jorge Matheu. Organización Panamericana de la Salud. Área de Vigilancia de la Salud y Prevención y Control de Enfermedades; Estados Unidos.Fil: Ramón Pardo, Pilar. Organización Panamericana de la Salud. Área de Vigilancia de la Salud y Prevención y Control de Enfermedades; Estados Unidos.Galas, Marcelo. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Bacteriología. Servicio Antimicrobianos; Argentina.OBJETIVO: Evaluar la capacidad de 17 laboratorios nacionales de referencia que participan en el Programa Latinoamericano de Control de Calidad en Bacteriología y Resistencia a los Antimicrobianos (LA-EQAS) para detectar mecanismos de resistencia emergentes, a saber: resistencia de enterobacterias a carbapenemes por presencia de Klebsiella pneumoniae carbapenemasa (KPC); resistencia de enterobacterias a carbapenemes por presencia de metalobetalactamasas (MBL) tipo IMP, y resistencia intermedia a vancomicina de aislamientos de Staphylococcus aureus (VISA). MÉTODOS: Se enviaron los siguientes tres aislamientos a los 17 laboratorios participantes del LA-EQAS: Klebsiella pneumoniae OPS-161 productor de KPC, Enterobacter cloacae OPS-166 productor de IMP y S. aureus OPS-165 con resistencia intermedia a vancomicina. Se evaluó la interpretación de las pruebas de sensibilidad y detección del mecanismo de resistencia y el tamaño de los halos de inhibición (método de difusión por discos) o valor de la concentración inhibitoria mínima (CIM). RESULTADOS: La concordancia en la detección de los mecanismos de resistencia fue de 76,4%, 73,3% y 66,7% con respecto a la cepas K. pneumoniae OPS-161, E. cloacae OPS-166 y S. aureus OPS-165, respectivamente. La concordancia entre las zonas de inhibición obtenidas por los laboratorios participantes y los rangos establecidos por el laboratorio coordinador fue aceptable en los tres aislamientos, ya que alcanzó 90,8%, 92,8% y 88,9%, respectivamente, para cada cepa. CONCLUSIONES: La concordancia global en la detección de los mecanismos de resistencia KPC, MBL y VISA fue de 72,1%. Consideramos que los laboratorios nacionales de referencia de América Latina son capaces de reconocer estos mecanismos de resistencia emergentes y se espera que en el futuro la concordancia alcance su nivel máximo

Population structure of Salmonella enterica Typhi in Harare, Zimbabwe (2012–19) before typhoid conjugate vaccine roll-out: A genomic epidemiology study

Background: The continued emergence of Salmonella enterica serovar Typhi, with ever increasing antimicrobial resistance, necessitates the use of vaccines in endemic countries. A typhoid fever outbreak in Harare, Zimbabwe, in 2018 from a multidrug resistant S Typhi with additional resistance to ciprofloxacin was the catalyst for the introduction of a typhoid conjugate vaccine programme. We aimed to investigate the emergence and evolution of antimicrobial resistance of endemic S Typhi in Zimbabwe and to determine the population structure, gene flux, and sequence polymorphisms of strains isolated before a typhoid conjugate vaccine programme to provide a baseline for future evaluation of the effect of the vaccination programme. Methods: In this genomic epidemiology study, we used short-read whole-genome sequencing of S Typhi isolated from clinical cases of typhoid fever in Harare, Zimbabwe, between Jan 1, 2012, and Feb 9, 2019, to determine the S Typhi population structure, gene flux, and sequence polymorphisms and reconstructed the evolution of antimicrobial resistance. Maximum likelihood time-scaled phylogenetic trees of Zimbabwe isolates in the context of global isolates obtained from the National Center for Biotechnology Information were constructed to infer spread and emergence of antimicrobial resistance. Findings: The population structure of S Typhi in Harare, Zimbabwe, from 2012 to 2019 was dominated by multidrug resistant genotype 4.3.1.1.EA1 (H58) that spread to Zimbabwe from neighbouring countries in around 2009 (95% credible interval 2008·5–2010·0). Acquisition of an IncN plasmid carrying antimicrobial resistance genes including a qnrS gene and a mutation in the quinolone resistance determining region of gyrA gene contributed to non-susceptibility and resistance to quinolone antibiotics. A minority population of antimicrobial susceptible S Typhi genotype 3.3.1 strains were present throughout. Interpretation: The currently dominant S Typhi population is genotype 4.3.1.1 that spread to Zimbabwe and acquired additional antimicrobial resistance though acquisition of a plasmid and mutation in the gyrA gene. This study provides a baseline population structure for future evaluation of the effect of the typhoid conjugate vaccine programme in Harare

mTOR inhibition decreases SOX2-SOX9 mediated glioma stem cell activity and temozolomide resistance

<p><b>Background</b>: SOX2 and SOX9 are commonly overexpressed in glioblastoma, and regulate the activity of glioma stem cells (GSCs). Their specific and overlapping roles in GSCs and glioma treatment remain unclear.</p> <p><b>Methods</b>: <i>SOX2</i> and <i>SOX9</i> levels were examined in human biopsies. Gain and loss of function determined the impact of altering SOX2 and SOX9 on cell proliferation, senescence, stem cell activity, tumorigenesis and chemoresistance.</p> <p><b>Results</b>: SOX2 and SOX9 expression correlates positively in glioma cells and glioblastoma biopsies. High levels of SOX2 bypass cellular senescence and promote resistance to temozolomide. Mechanistic investigations revealed that SOX2 acts upstream of SOX9. mTOR genetic and pharmacologic (rapamycin) inhibition decreased SOX2 and SOX9 expression, and reversed chemoresistance.</p> <p><b>Conclusions</b>: Our findings reveal SOX2-SOX9 as an oncogenic axis that regulates stem cell properties and chemoresistance. We identify that rapamycin abrogate SOX protein expression and provide evidence that a combination of rapamycin and temozolomide inhibits tumor growth in cells with high SOX2/SOX9.</p