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...

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.

Búsqueda activa de individuos con tuberculosis pulmonar y extrapulmonar en Calarcá-Quindío, Colombia-2005 / Detecting active tuberculosis in Calarcá-Quindío, Colombia, during 2005

Objetivo Describir características socio demográficas en una muestra de 195 pacientes sintomáticos respiratorios y 18 pacientes tuberculosos de Calarcá - Quindío, Colombia en el 2005. Metodología Se realizó búsqueda activa de tuberculosos en tres zonas urbanas, la cárcel del municipio de Calarcá y el Hospital Universitario San Juan de Dios. Previo ingreso al estudio los pacientes diligenciaron una encuesta y un consentimiento informado. A cada paciente se le tomaron muestras de esputo para baciloscopia y cultivo en medio de Ogawa-Kudoh. Cuando se observó crecimiento se realizó identificación fenotípica por pruebas bioquímicas y genotipificación por spoligotyping. Resultados Se identificaron 195 pacientes sintomáticos respiratorios y 18 (9,2 por ciento) pacientes tuberculosos, de ellos el 78 por ciento corresponde al género masculino y 22 por ciento al femenino con una edad promedio para ambos sexos de 40 años. Se procesaron un total de 174 muestras de esputo, 21 lavados bronco alveolares, una muestra de orina, una muestra de ganglio cervical y un liquido pleural. El 77 por ciento de las muestras analizadas fueron positivos por examen directo y cultivo, 16,7 por ciento por cultivo y el 5,5 por ciento por baciloscopia únicamente. El promedio de baciloscopias por paciente fue 2,3. La prueba de spoligotyping realizada evidenció la presencia de los genotipos H1, T1, H3 y LAM9. Conclusión Los resultados obtenidos demuestran la importancia de realizar simultáneamente baciloscopia y cultivo para aumentar la detección de pacientes bacilíferos sintomáticos respiratorios, al igual que en el seguimiento y control de estos pacientes. Se demostró la presencia de cuatro genotipos diferentes de Mycobacterium tuberculosis en Calarcá.

Objective Describing socio-demographic characteristics in a population of 195 respiratory symptomatic and 18 tuberculosis patients from Calarcá-Quindío, Colombia during 2005. Methods An active search for individuals having symptomatic tuberculosis was carried out in three urban areas, Calarca's municipal prison and the San Juan de Dios teaching hospital. Prior to entering the study, the patients were required to fill out an application form and sign an informed consent form. Sputum samples were taken from each patient for bacilloscopy and culture in Ogawa-Kudoh medium. Phenotypic and genotypic identification were made when growth was observed by using conventional and molecular methods. Results 18 (9 ,2 percent) out of 195 individuals having respiratory symptoms were diagnosed as having tuberculosis. 78 percent were male and 22 percent female; the average age was 40 for both genders. 174 sputum, 21 bronchial lavage, one urine, one cervical ganglia and pleural liquid sample were analysed; 77 percent proved positive by direct smear test and culture, 16,7 percent by culture and only 5,5 percent by direct smear test. Bacilloscopy average per patient was 2 ,3 . The presence of H1, T1, H3 and LAM9 genotypes was demonstrated by using the spoligotyping test. Conclusions The results confirmed the importance of using both direct smear test and/or culture for improving the early detection of pulmonary tuberculosis, its follow-up and control. The existence of four different Mycobacterium tuberculosis genotypes in Calarcá was established.