Metagenomic and genomic analysis of heavy metal-tolerant and -resistant bacteria in resource islands in a semi-arid zone of the Colombian Caribbean.

Bacteria from resource islands can adapt to different extreme conditions in semi-arid regions. We aimed to determine the potential resistance and tolerance to heavy metals from the bacterial community under the canopy of three resource islands in a semi-arid zone of the Colombian Caribbean. Total DNA was extracted from soil and through a metagenomics approach, we identified genes related to heavy metal tolerance and resistance under the influence of drought and humidity conditions, as well as the presence or absence of vegetation. We characterized the genomes of bacterial isolates cultivated in the presence of four heavy metals. The abundances of genes related to heavy metal resistance and tolerance were favored by soil moisture and the presence of vegetation. We observed a high abundance of resistance genes (60.4%) for Cu, Zn, and Ni, while 39.6% represented tolerance. These genes positively correlated with clay and silt content, and negatively correlated with sand content. Resistance and tolerance were associated with detoxification mechanisms involving oxidoreductase enzymes, metalloproteases, and hydrolases, as well as transmembrane proteins involved in metal transport such as efflux pumps and ion transmembrane transporters. The Bacillus velezensis C3-3 and Cytobacillus gottheilii T106 isolates showed resistance to 5 mM of Cd, Co, Mn, and Ni through detoxification genes associated with ABC pumps, metal transport proteins, ion antiporter proteins, and import systems, among others. Overall, these findings highlight the potential of bacteria from resource islands in bioremediation processes of soils contaminated with heavy metals.

Bioinformatic approach for repurposing immunomodulatory drugs for lepromatous leprosy.

Background: The lepromatous leprosy (LL) disease is caused by Mycobacterium leprae and Mycobacterium lepromatosis which is characterized by inadequate response to treatment, a propensity to drug resistance, and patient disability. We aimed to evaluate current immunomodulatory medicines and their target proteins collectively as a drug repurposing strategy to decipher novel uses for LL. Methods: A dataset of human genes associated with LL-immune response was retrieved from public health genomic databases including the Human Genome Epidemiology Navigator and DisGeNET. Retrieved genes were filtered and enriched to set a robust network (≥10, up to 21 edges) and analyzed in the Cytoscape program (v3.9). Drug associations were obtained in the NDEx Integrated Query (v1.3.1) coupled with drug databases such as ChEMBL, BioGRID, and DrugBank. These networks were analyzed in Cytoscape with the CyNDEx-2 plugin and STRING protein network database. Results: Pathways analyses resulted in 100 candidate drugs organized into pharmacological groups with similar targets and filtered on 54 different drugs. Gene-target network analysis showed that the main druggable targets associated with LL were tumoral necrosis factor-alpha, interleukin-1B, and interferon-gamma. Consistently, glucosamine, binimetinib, talmapimod, dilmapimod, andrographolide, and VX-702 might have a possible beneficial effect coupled with LL treatment. Conclusion: Based on our drug repurposing analysis, immunomodulatory drugs might have a promising potential to be explored further as therapeutic options or to alleviate symptoms in LL patients.

Regulations on the Use of Antibiotics in Livestock Production in South America: A Comparative Literature Analysis.

As a global health problem, antimicrobial resistance (AMR) crosses national borders, leading UN (United Nations) multilateral agencies to call for all countries to improve the stewardship of antibiotics in humans and animals. South American countries have changed their regulations regarding antibiotic use in livestock production. This literature review examines how far the five largest meat-producing countries in South America (Argentina, Brazil, Chile, Colombia, and Uruguay) have come in terms of the relevant legislation. Rules on market entry (marketing authorization and official distribution systems) are already set in all countries examined. Four countries do not allow growth promoters based on critically important antibiotics, and countries have also begun to set guidelines and minimum welfare and biosecurity requirements to reduce the therapeutic demand for antibiotics. Nonetheless, there are aspects related to the distribution, use, and disposal of antibiotics that need to be developed further. In conclusion, legislation in South American countries is moving towards the goals set by UN multilateral agencies, but more can be done. Differences between countries' rules and the gold standards set by the World Organization for Animal Health (OIE), World Health Organization (WHO), and Food and Agriculture Organization of the United Nations (FAO) reveal possible adaptations to the countries' realities. Further studies must examine compliance with the legislation already set and investigate other tools that can be used alongside legislation as a driving force to change stakeholder behaviour.

Specific identification of Mycobacterium bovis by Loop-Mediated Isothermal Amplification (LAMP) targeting the Region of Difference 12 (RD12) of the M. tuberculosis complex.

Bovine tuberculosis is a prevalent zoonotic disease that causes high risks for production animals, dairy producers and consumers, together with significant economic losses. Thus, methods for easy, fast and specific detection of Mycobacterium bovis in small and medium-sized livestock under field conditions are very required. In this work, a Loop-Mediated Isothermal Amplification LAMP-PCR targeting the Region of Difference 12 (RD12) of M. bovis genome was designed for the purpose of identification. A set of six primers designed for the isothermal amplification of five different genomic fragments led to the specific identification of M. bovis from other mycobacterial species. A basic colorimetric reaction was clearly observed at first sight under natural light, indicating positive identification of M. bovis in a maximum of 30 min of isothermal amplification at 65 °C. The limit of detection was near 50 fg of M. bovis genomic DNA, corresponding approximately to 10 copies of the genome. •The proposed LAMP-PCR amplification of M. bovis genomic DNA might be performed by untrained laboratory personnel.•Specific identification of M. bovis LAMP is possible in 30 min at 65.. C using a simple water bath.•The basic colorimetric reaction for M. bovis identification could be observed with the naked eye under natural light.

Design of a specific peptide against phenolic glycolipid-1 from Mycobacterium leprae and its implications in leprosy bacilli entry.

BACKGROUND: Mycobacterium leprae, the causative agent of Hansen's disease, causes neural damage through the specific interaction between the external phenolic glycolipid-1 (PGL-1) and laminin subunit alpha-2 (LAMA2) from Schwann cells. OBJECTIVE: To design a LAMA2-based peptide that targets PGL-1 from M. leprae. METHODS: We retrieved the protein sequence of human LAMA2 and designed a specific peptide using the Antimicrobial Peptide Database and physicochemical parameters for antimycobacterial peptide-lipid interactions. We used the AlphaFold2 server to predict its three-dimensional structure, AUTODOCK-VINA for docking, and GROMACS programs for molecular dynamics simulations. FINDINGS: We analysed 52 candidate peptides from LAMA2, and subsequent screening resulted in a single 60-mer peptide. The mapped peptide comprises four ß-sheets and a random coiled region. This peptide exhibits a 45% hydrophobic ratio, in which one-third covers the same surface. Molecular dynamics simulations show that our predicted peptide is stable in aqueous solution and remains stable upon interaction with PGL-1 binding. In addition, we found that PGL-1 has a preference for one of the two faces of the predicted peptide, which could act as the preferential binding site of PGL-1. MAIN CONCLUSIONS: Our LAMA2-based peptide targeting PGL-1 might have the potential to specifically block this key molecule, suggesting that the preferential region of the peptide is involved in the initial contact during the attachment of leprosy bacilli to Schwann cells.

Design of a specific peptide against phenolic glycolipid-1 from Mycobacterium leprae and its implications in leprosy bacilli entry

BACKGROUND Mycobacterium leprae, the causative agent of Hansen's disease, causes neural damage through the specific interaction between the external phenolic glycolipid-1 (PGL-1) and laminin subunit alpha-2 (LAMA2) from Schwann cells. OBJECTIVE To design a LAMA2-based peptide that targets PGL-1 from M. leprae. METHODS We retrieved the protein sequence of human LAMA2 and designed a specific peptide using the Antimicrobial Peptide Database and physicochemical parameters for antimycobacterial peptide-lipid interactions. We used the AlphaFold2 server to predict its three-dimensional structure, AUTODOCK-VINA for docking, and GROMACS programs for molecular dynamics simulations. FINDINGS We analysed 52 candidate peptides from LAMA2, and subsequent screening resulted in a single 60-mer peptide. The mapped peptide comprises four β-sheets and a random coiled region. This peptide exhibits a 45% hydrophobic ratio, in which one-third covers the same surface. Molecular dynamics simulations show that our predicted peptide is stable in aqueous solution and remains stable upon interaction with PGL-1 binding. In addition, we found that PGL-1 has a preference for one of the two faces of the predicted peptide, which could act as the preferential binding site of PGL-1. MAIN CONCLUSIONS Our LAMA2-based peptide targeting PGL-1 might have the potential to specifically block this key molecule, suggesting that the preferential region of the peptide is involved in the initial contact during the attachment of leprosy bacilli to Schwann cells.

Interactive web-based tool for evaluating the spread of bovine tuberculosis and brucellosis in Colombia / Herramienta interactiva basada en la web para evaluar la propagación de la tuberculosis bovina y la brucelosis en Colombia / Ferramenta interativa baseada na web para avaliar a disseminação da tuberculose e brucelose bovina na Colômbia

Abstract Background: Bovine tuberculosis (BTB) and brucellosis are associated with devastating losses in the livestock sector in Colombia and even in developed countries. Real-time disease surveillance is a key strategy to control and eradicate infectious disease outbreaks. Objective: To design an epidemiological tool for monitoring BTB and brucellosis in Colombia. Methods: An interactive platform for disease mapping of BTB and brucellosis during an observation period between years 2004 and 2019 was designed. Results: Our analysis showed that the provinces of Cundinamarca and Valle del Cauca are regions affected by BTB and brucellosis epidemics, respectively (p<0.001). Furthermore, increased case detection of BTB was reported in 2012 and brucellosis in 2019 (p<0.001). Conclusions: This epidemiological platform allows tracking BTB and tuberculosis hotspots, identifying trends over time, and provides useful information to animal health authorities for designing new strategies in control programs.

Resumen Antecedentes: La tuberculosis bovina (TBB) y la brucelosis están asociadas con problemas persistentes en la ganadería Colombiana e incluso en los países desarrollados. La vigilancia de enfermedades en tiempo real es una estrategia clave para controlar y erradicar brotes de enfermedades infecciosas. Objetivo: Diseñar una herramienta epidemiológica para monitorear TBB y brucelosis en Colombia. Métodos: Se diseñó un panel de control interactivo para el mapeo de ambas enfermedades durante el periodo de observación entre los años 2004 y 2019. Resultados: El análisis de la herramienta mostró que las Provincias de Cundinamarca y Valle del Cauca han sido áreas epidémicas para TBB y brucelosis, respectivamente (p<0,001). Además, se encontró un aumento de la detección de casos de TBB en 2012 y de brucelosis durante 2019 (p<0,001). Conclusiones: Este panel epidemiológico permite el seguimiento de puntos críticos de TBB y tuberculosis, identificando sus tendencias a lo largo del tiempo, y proporciona información útil para las autoridades de sanidad animal que diseñan nuevas estrategias para los programas de control.

Resumo Antecedentes: A tuberculose bovina (TBB) e a brucelose estão associadas a problemas persistentes no campo da pecuária na Colômbia e até em países desenvolvidos. Portanto, a vigilância de doenças em tempo real é uma estratégia essencial para controlar e erradicar surtos de doenças infecciosas. Objetivo: Projetar uma ferramenta epidemiológica para monitorar a TB e a brucelose na Colômbia. Métodos: Um painel de controle interativo foi projetado para o mapeamento de ambas as doenças entre 2004 e 2019 como período de observação. Resultados: A análise da ferramenta mostrou que as Províncias de Cundinamarca e Valle del Cauca foram áreas epidêmicas para TBB e brucelose, respectivamente (p<0,001). Além disso, foi encontrado um aumento na detecção de casos em 2012 para TBB e brucelose durante 2019 (p<0,001). Conclusões: Esse painel epidemiológico poderia permitir o monitoramento de pontos críticos dessas doenças, identificando tendências ao longo do tempo, fornecendo informações úteis para as autoridades de saúde animal que elaboram novas estratégias para programas de controle.

Characterising four Sarconesiopsis magellanica (Diptera: Calliphoridae) larval fat body-derived antimicrobial peptides.

BACKGROUND: The inappropriate use of antibiotics has led to the accelerated growth of resistance to antibiotics. The search for new therapeutic strategies (i.e., antimicrobial peptides-AMPs) has thus become a pressing need. OBJECTIVE: Characterising and evaluating Sarconesiopsis magellanica larval fat body-derived AMPs. METHODS: Fat body extracts were analysed by reversed-phase high-performance liquid chromatography (RP-HPLC); mass spectrometry was used for characterising the primary structure of the AMPs so found. ProtParam (Expasy) was used for analysing the AMPs' physico-chemical properties. Synthetic AMPs' antibacterial activity was evaluated. FINDINGS: Four new AMPs were obtained and called sarconesin III, IV, V and VI. Sarconesin III had an α-helix structure and sarconesins IV, V and VI had linear formations. Oligomer prediction highlighted peptide-peptide interactions, suggesting that sarconesins III, V and VI could form self-aggregations when in contact with the microbial membrane. AMPs synthesised from their native molecules' sequences had potent activity against Gram-positive bacteria and, to a lesser extent, against Gram-negative and drug-resistant bacteria. Sarconesin VI was the most efficient AMP. None of the four synthetic AMPs had a cytotoxic effect. MAIN CONCLUSIONS: S. magellanica larval fat body-derived antimicrobial peptides are an important source of AMPs and could be used in different antimicrobial therapies and overcoming bacterial resistance.

Genes associated with antibiotic tolerance and synthesis of antimicrobial compounds in a mangrove with contrasting salinities.

Salinity and wastewater pollution in mangrove ecosystems can affect microorganisms and the abundance of genes involved in response to these stressors. This research aimed to identify genes associated with resistance and biosynthesis of antimicrobial compounds in mangrove soils subjected to contrasting salinities and wastewater pollution. Samples of rhizospheric soil were taken from a mangrove at the mouth of the Ranchería River in La Guajira, Colombia. A functional analysis was performed using Illumina HiSeq 2500 sequencing data obtained from total DNA extracted. Increased salt concentration influenced metabolic pathways and differential abundance of genes associated with the synthesis of antimicrobial compounds (e.g., rfbB/rffG, INO1/ISYNA1, rfbA/rffH, sat/met3, asd). Also, among 33 genes involved in intrinsic antibiotic resistance, 16 were significantly influenced by salinity (e.g., cusR/copR/silR, vgb, tolC). We concluded that salt stress tolerance and adaptive mechanisms could favor the biosynthesis of antimicrobial compounds in mangroves contaminated by sewage.

Characterising four Sarconesiopsis magellanica (Diptera: Calliphoridae) larval fat body-derived antimicrobial peptides

BACKGROUND The inappropriate use of antibiotics has led to the accelerated growth of resistance to antibiotics. The search for new therapeutic strategies (i.e., antimicrobial peptides-AMPs) has thus become a pressing need. OBJECTIVE Characterising and evaluating Sarconesiopsis magellanica larval fat body-derived AMPs. METHODS Fat body extracts were analysed by reversed-phase high-performance liquid chromatography (RP-HPLC); mass spectrometry was used for characterising the primary structure of the AMPs so found. ProtParam (Expasy) was used for analysing the AMPs' physico-chemical properties. Synthetic AMPs' antibacterial activity was evaluated. FINDINGS Four new AMPs were obtained and called sarconesin III, IV, V and VI. Sarconesin III had an α-helix structure and sarconesins IV, V and VI had linear formations. Oligomer prediction highlighted peptide-peptide interactions, suggesting that sarconesins III, V and VI could form self-aggregations when in contact with the microbial membrane. AMPs synthesised from their native molecules' sequences had potent activity against Gram-positive bacteria and, to a lesser extent, against Gram-negative and drug-resistant bacteria. Sarconesin VI was the most efficient AMP. None of the four synthetic AMPs had a cytotoxic effect. MAIN CONCLUSIONS S. magellanica larval fat body-derived antimicrobial peptides are an important source of AMPs and could be used in different antimicrobial therapies and overcoming bacterial resistance.

Characterising four Sarconesiopsis magellanica (Diptera: Calliphoridae) larval fat body-derived antimicrobial peptides

BACKGROUND The inappropriate use of antibiotics has led to the accelerated growth of resistance to antibiotics. The search for new therapeutic strategies (i.e., antimicrobial peptides-AMPs) has thus become a pressing need. OBJECTIVE Characterising and evaluating Sarconesiopsis magellanica larval fat body-derived AMPs. METHODS Fat body extracts were analysed by reversed-phase high-performance liquid chromatography (RP-HPLC); mass spectrometry was used for characterising the primary structure of the AMPs so found. ProtParam (Expasy) was used for analysing the AMPs’ physico-chemical properties. Synthetic AMPs’ antibacterial activity was evaluated. FINDINGS Four new AMPs were obtained and called sarconesin III, IV, V and VI. Sarconesin III had an α-helix structure and sarconesins IV, V and VI had linear formations. Oligomer prediction highlighted peptide-peptide interactions, suggesting that sarconesins III, V and VI could form self-aggregations when in contact with the microbial membrane. AMPs synthesised from their native molecules’ sequences had potent activity against Gram-positive bacteria and, to a lesser extent, against Gram-negative and drug-resistant bacteria. Sarconesin VI was the most efficient AMP. None of the four synthetic AMPs had a cytotoxic effect. MAIN CONCLUSIONS S. magellanica larval fat body-derived antimicrobial peptides are an important source of AMPs and could be used in different antimicrobial therapies and overcoming bacterial resistance

Producción pecuaria y emergencia de antibiótico resistencia en Colombia: Revisión sistemática / Livestock production and emergency antibiotic resistance in Colombia: Systematic review

El uso extensivo de antibióticos es una práctica comúnmente realizada para aumentar la producción pecuaria. Así, la crianza animal demanda una fuerte presión selectiva para prevenir brotes de infecciones pero que también podría resultar en la emergencia de cepas multidrogoresistentes. El propósito de esta revisión, es documentar la posible contribución de las practicas pecuarias en la emergencia de patógenos zoonóticos que exhiben resistencia a antibióticos en Colombia. Los patógenos resistentes a antibióticos reportados con mayor frecuencia y asociados a alimentos fueron Salmonella sp. y Escherichia coli. Se encontró que el uso no terapéutico y abuso de antibióticos β-lactámicos, macrólidos y tetraciclinas constituyen la mayor presión selectiva. Adicionalmente, se encontraron estudios locales que reportan la contaminación de fuentes ambientales y alimentos con trazas de antibióticos. La aparición de patógenos resistentes a antibióticos de uso veterinario podría ser producto de la precaria implementación de buenas prácticas pecuarias entorno al componente de sanidad animal.

The extensive use of antibiotics is a common practice to increase livestock production. Thus, animal husbandry entails a high selective pressure to control infectious outbreaks which also might result in the emergence of multidrug resistant strains. This review's aim is to survey cases associated with zoonotic pathogens showing antibiotic resistance in Colombia. The resistant pathogens most commonly isolated from the food chain were Salmonella sp. and Escherichia coli. The nontherapeutic and abuse of antibiotics such as β-lactams, macrolides and tetracycline represented the most critical selective pressure. Furthermore, environmental and food contamination with traces of antibiotics have been found in different local studies. Rise of resistant pathogens to veterinary drugs might result due poor implementation of good farming practices in the animal health plan.

Complete Genome Sequence of Escherichia coli AS19, an Antibiotic-Sensitive Variant of E. coli Strain B REL606.

The chemically mutagenized Escherichia coli strain AS19 was isolated on the basis of its enhanced sensitivity to different antibiotics, in particular to actinomycin. The strain was later modified to study rRNA modifications that confer antibiotic resistance. Here, we present the genome sequence of the variant E. coli AS19-RrmA.

Caracterización socio-económica y ambiental en pequeños y medianos predios ganaderos en la región del sumapaz, colombia

Las comunidades ganaderas, a pequeña y mediana escala, en la provincia del Sumapaz, se encuentran en un periodo de transición hacia la tecnificación en la producción. Así, el objetivo de este estudio fue describir algunos aspectos que actúan como barreras en la tecnificación y limitan la implementación de buenas prácticas ganaderas. Se realizó un diagnóstico participativo en pequeños y medianos ganaderos del Sumapaz, mediante encuesta aleatoria, en nueve municipios de la provincia. Los resultados evidencian que existen diferentes barreras socio-económicas, que obstaculizan el acceso a tecnologías para el mejoramiento de la producción, como el bajo nivel de ingresos económicos, acceso a la educación superior y falta de registros contables y de sanidad en la producción; sin embargo, se observaron indicadores positivos, asociados al manejo ambiental de los recursos naturales, como parte de su legado cultural.

Livestock community from the Sumapaz region are in a transition period towards to technical production. Thus, our aim was to assess the mean requirements which delay implementation of good farming practices. This study, involved small and medium livestock farmers from the Sumapaz region through a random test applied in nine municipalities from the province. Results showed socioeconomic barriers which limited the technology transfer for improvement in production such as low income, restricted access to university education and lack of productive and sanitary records. However, we observed positive outcomes with regard to environmental management which is characteristic as their cultural heritage.

Screening food-borne and zoonotic pathogens associated with livestock practices in the Sumapaz region, Cundinamarca, Colombia.

Hazardous practices regarding antibiotics misuse, unsanitary milking procedures, and the commercial sales of raw milk and unpasteurized dairy products are currently being practiced by livestock farmers in the Sumapaz region (Colombia). The purpose of this study was to screen for food-borne and zoonotic pathogens associated with local livestock practices. We evaluated 1098 cows from 46 livestock farms in the Sumapaz region that were selected by random. Of the total population of cattle, 962 animals (88%) were tested for bovine TB using a caudal-fold tuberculin test and 546 (50%) for brucellosis by a competitive ELISA. In the population tested, 23 cows were positive for Brucella sp. representing a 4.2% seroprevalence and no cases of bovine tuberculosis were found. In addition, food-borne contamination with Escherichia coli and Staphylococcus aureus was assessed together with antibiotic susceptibility for ten different antibiotics in milk samples from 16 livestock farms. We found that 12 of the farms (75%) were contaminated with these food-borne pathogens. Noteworthy, all of the isolated pathogenic strains were resistant to multiple antibiotics, primarily to oxytetracycline and erythromycin. Our findings suggest that livestock products could be a source of exposure to Brucella and multidrug-resistant E. coli and S. aureus strains as a result of unhygienic livestock practices in the Sumapaz region. Training in good farming practices is the key to improving safety in food production.

Structural features of the two-component systemLisR/LisK suggests multiple responses for the adaptation and survival of Listeria monocytogenes

Se caracterizó la estructura del sistema deregulación de dos componentes LisR/LisK de Listeriamonocytogenes. Se emplearon herramientas bioinformáticas ybases de datos para predecir la estructura e interacciones delas dos proteínas y se modelaron. Los resultados predicenque la proteína LisK está embebida en la membrana celulary su composición modular (dominios HAMP, histidinakinasa and ATPasa) está asociada a su autofosforilación(His-266). Un efecto estímulo respuesta determina lapropagación secuencial de la señal desde la membranacelular hacia componentes citoplasmáticos. A su vez, sepredice que LisR es una proteína citosólica con un dominiode receptor (homólogo a cheY) que incluye el residuofosfo-aceptor (Asp-52) y el dominio de unión a ADN,el cual puede permitir la transmisión de una respuestaespecífica a nivel transcripcional. Los componentes LisR/LisK han sido bioquímica y funcionalmente caracterizadosexperimentalmente en la patofisiología de otros bacilos. Espor ello, que la aproximación de los resultados basados enestructura-función podría facilitar el diseño de inhibidoresespecíficos...

Here, we characterized the structure of the two-component regulatory system, LisR/LisK, in Listeriamonocytogenes. To predict the structure of both proteins and the relationship between them, we employedseveral bioinformatic tools and databases. Based on our results, LisK protein is embedded in the cellmembrane and its modular composition (HAMP, histidine kinase and ATPase domains) is associatedwith its autophosphorylation (His-266). A stimulus-response likely determines the sequential signalpropagation from the bacterial cell surface to its cytoplasmic components. According to our results,LisR is a cytoplasmic protein with a receptor domain (homologous to CheY) that comprises a phosphoacceptorresidue (Asp-52) and a DNA-binding domain, which may allow the transmission of a specifictranscriptional response. LisR/LisK has been experimentally characterized both biochemically andfunctionally in other Bacilli pathophysiology; our structure-function approach may facilitate the design ofsuitable inhibitors...

O objetivo do estudo foi caracterizarestruturalmente o sistema de regulação de dois componentesLisR/LisK de Listeria monocytogenes. Foram utilizadasdiversas ferramentas de bioinformática e bancos de dadospara predizer a estrutura das duas proteínas, modelálase prever suas interações. Os resultados predizemque a proteína Lisk está incorporada na composição damembrana celular e sua composição modular (domíniosHAMP, histidina quinase e ATPase) está associada com asua autofosforilação (His-266). Um efeito de estímulo eresposta determina a propagação sequencial do sinal a partirda membrana celular em componentes citoplasmáticos. Osresultados predizem que LisR é uma proteína citosólicacom um domínio recetor (homólogo a CheY) que inclui oresíduo fosfo-aceitador (Asp-52) e o domínio de ligação aoADN, o que pode permitir a transmissão de uma respostaespecífica a nível transcricional. Como LisR/Lisk foi,química e funcionalmente, caracterizada experimentalmentena fisiopatologia de outros bacilos, esta abordagem baseadana estrutura-função pode facilitar a conceção de inibidoresespecíficos...

Estado de la coinfección tuberculosis/virus de la inmunodeficiencia humana en el municipio de Armenia(Colombia): experiencia de 10 años / Status of tuberculosis/HIV coinfection in the municipality of Armenia (Colombia): Ten years experience

La tuberculosis (TB) representa una de las principales complicaciones clínicas en personas inmunocomprometidas por virus de la inmunodeficiencia humana (VIH). Así, cuando las acciones diagnósticas y terapéuticas no son tempranas, el resultado puede llegar a ser letal. Metodología: Se llevó a cabo un estudio descriptivo y retrospectivo de los casos de coinfección TB/VIH en el período comprendido entre los años 2000 y 2010 en el municipio de Armenia. Se revisaron las fichas de notificación de ambos eventos (Sistema Nacional de Vigilancia en Salud Pública), actas de visitas domiciliarias, tarjetas individuales de tratamiento y certificados de defunción. Resultados: Se incluyeron 113 pacientes con diagnóstico confirmado de coinfección TB/VIH, lo que representa una prevalencia global de 6,8% en 10 años en Armenia. La edad promedio de los pacientes fue 34,3 años (11 a 68 años), y los más comprometidos fueron los del grupo de 14 a 34 años de sexo masculino (91% en formas extrapulmonares y 75% en pulmonares). La TB extrapulmonar o la miliar predominaron con un 50,4% (n = 57), sobre la TB pulmonar. Las formas extrapulmonares más comunes fueron: ganglionar 50% (n = 32), meníngea 19,3% (n = 11), miliar 8,8% (n = 5), pleural 7% (n = 4), peritoneal 7% (n = 4) y vertebral (mal de Pott) 1,8% (n = 1). En 8 casos hubo presentación mixta (pulmonar y extrapulmonar). La letalidad fue del 100% en pacientes infectados por VIH y asociados con cepas resistentes. Conclusiones: Los resultados de este estudio resaltan la importancia de implementar acciones colaborativas entre los programas de TB/VIH para mejorar el diagnóstico y reducir la mortalidad la diseminación de cepas resistentes.

Tuberculosis represents one of the top clinical complications in immunocompromised HIVinfected people. When diagnostic and therapeutic interventions are delayed, outcomes could be fatal due to imminent progress of this deadly combination. Methodology: We carried out a descriptive retrospective study from registered TB/HIV cases from 2000 to 2010 in the municipality of Armenia. Notification forms from both diseases TB and HIV (SIVIGILA), record of home visits, individual treatment cards and death certificates were used as data sources. Results: 113 patients were included with TB/HIV confirmed tests, which represents a global prevalence of 6.8% during 10 years in Armenia. The mean age among patients was 34.3 years (ranging from 11 to 68 years), and the group between 14 and 34 years and male (91% extrapulmonary and 75% pulmonary forms) was the most commonly involved. According to the disease presentation, extrapulmonary TB form (50.4%, n = 57) was more frequent than the pulmonary form. The frequency of extrapulmonary forms was: ganglionar, 50% (n = 32); meningeal, 19.3% (n = 11); miliar, 8.8% (n = 5); pleural, 7% (n = 4); peritoneal, 7% (n = 4), and Pott’s disease, 1.8% (n = 1). In eight cases both forms presented simultaneously (pulmonary and extrapulmonary). Lethality was 100% among VIH positive patients infected with resistant strains. Conclusions: Our results highlight the importance of implementing collaborative actions between TB and HIV programs to improve diagnosis and to reduce mortality and resistant strain dissemination.

Análisis sociodemográfico y espacial de la transmisión de la tuberculosis en la ciudad de Armenia (Colombia) / Sociodemographic and spatial transmission of tuberculosis in the city of Armenia (Colombia)

La tuberculosis es uno de los problemas que demanda atención prioritaria en el municipio de Armenia, capital del departamento del Quindío (Colombia). En este estudio, se encontró que la población afectada, generalmente pertenece al género masculino (razón hombre: mujer 2:1) y en edad económicamente productiva (25-54 años). La mayoría de enfermos son del régimen subsidiado (40%) o no asegurados (26%) y viven en condiciones socioeconómicas críticas. Se encontraron diferencias significativas de carga de enfermedad en las tasas entre comunas: en unas localidades fueron de 10 y en otras de 30 casos por 100.000 habitantes, indicando heterogeneidad de la situación en la ciudad. El análisis geoespacial permite caracterizar y precisar los escenarios donde se deben concentrar las acciones prioritarias del programa de control de la tuberculosis.

Tuberculosis is an important public health problem that demands priority action in the city of Armenia (Colombia). In this study, we found that most cases of TB were males (ratio 2:1) and economically active population (25-54 years). TB patients had subvention health care (40%) or were uncovered by the national health insurance (26%), living in critical socioeconomic condi tions. Significant differences in the TB prevalence rate between town localities were found; in some zones were 10 and 30 cases per 100.000 inhabitants, indicating a heterogeneous situation. Spatial analysis might allow the definition and concentration of early actions in a region for the tuberculosis control.

Características clínicas y sociodemográficas de los casos con tuberculosis resistente en el municipio de Armenia, Quindío (Colombia) / Clinical and socio-demographic characteristics of tuberculosis with antimicrobial resistance in Armenia, Quindio (Colombia)

Objetivo: Analizar los factores sociodemográficos y clínicos de los pacientes con diagnóstico de tuberculosis resistente en el municipio de Armenia durante el período 2006-2009. Métodos: Se realizó un estudio descriptivo y retrospectivo con base en la notificación de pacientes con tuberculosis resistentes durante el período de estudio. Los datos fueron tomados de los registros consignados en las fichas de notificación y las tarjetas individuales de tratamiento categoría IV. Resultados: Entre 2006 y 2009, 678 pacientes ingresaron en el programa de control de la tuberculosis. De ellos, 14 casos (2,0%) fueron resistentes al menos a un medicamento, y entre estos, 7 (50%) fueron multirresistentes. El 21% estuvo asociado a infección por virus de la inmunodeficiencia humana. La tasa de fallecimientos en pacientes con tuberculosis con farmacorresistencia múltiple fue del 71%. Se encontró amplificación de la resistencia en 2 casos, quienes presentaron deterioro en el estado de salud y deceso. Conclusión: La situación en Armenia de resistencia a antituberculosos es de proporción similar a la del resto del país. Se encontraron factores de riesgo conocidos en los casos de resistencia (coinfección con virus de la inmunodeficiencia humana, farmacodependencia, abandono social) en la mayoría de casos, pero también de resistencia primaria y sin factores de riesgo con farmacorresistencia múltiple y una alta mortalidad, lo que llama la atención para mejorar la vigilancia y el control de los casos en tratamiento en la ciudad.

Objective: To analyze the socio-demographic and clinical diagnosis of patients with resistant tuberculosis in the municipality of Armenia in the period 2006-2009. Methods: We performed a retrospective descriptive study based on the notification of patients with resistant tuberculosis during the study period, data were taken from registrations under notification forms and individual treatment cards category IV. Results: 678 patients were admitted to the program of tuberculosis control between 2006 and 2009, 14 cases (2,0%) were resistant to at least one drug and from these cases 7 were multidrug resistant (MDR). In 21% of cases there was co-infection with HIV. The death rate in patients with MDR-TB was 71%. Amplification in resistance was observed in two cases which lead to further deterioration in the health status and death. Conclusion: Armenia has levels of resistance to anti tuberculosis drugs similar to the reported in the rest of Colombia. We found factors already know to be associated with resistance (drug addiction, social abandon, HIV co infection) but there were also cases without these factors and with primary MDR and high mortality, therefore is urgent to improve the public health measures for cases under treatment in this city.

Molecular modeling and in silico characterization of Mycobacterium tuberculosis TlyA: possible misannotation of this tubercle bacilli-hemolysin.

BACKGROUND: The TlyA protein has a controversial function as a virulence factor in Mycobacterium tuberculosis (M. tuberculosis). At present, its dual activity as hemolysin and RNA methyltransferase in M. tuberculosis has been indirectly proposed based on in vitro results. There is no evidence however for TlyA relevance in the survival of tubercle bacilli inside host cells or whether both activities are functionally linked. A thorough analysis of structure prediction for this mycobacterial protein in this study shows the need for reevaluating TlyA's function in virulence. RESULTS: Bioinformatics analysis of TlyA identified a ribosomal protein binding domain (S4 domain), located between residues 5 and 68 as well as an FtsJ-like methyltranferase domain encompassing residues 62 and 247, all of which have been previously described in translation machinery-associated proteins. Subcellular localization prediction showed that TlyA lacks a signal peptide and its hydrophobicity profile showed no evidence of transmembrane helices. These findings suggested that it may not be attached to the membrane, which is consistent with a cytoplasmic localization. Three-dimensional modeling of TlyA showed a consensus structure, having a common core formed by a six-stranded ß-sheet between two α-helix layers, which is consistent with an RNA methyltransferase structure. Phylogenetic analyses showed high conservation of the tlyA gene among Mycobacterium species. Additionally, the nucleotide substitution rates suggested purifying selection during tlyA gene evolution and the absence of a common ancestor between TlyA proteins and bacterial pore-forming proteins. CONCLUSION: Altogether, our manual in silico curation suggested that TlyA is involved in ribosomal biogenesis and that there is a functional annotation error regarding this protein family in several microbial and plant genomes, including the M. tuberculosis genome.