RESUMO
Carrion's disease is a neglected endemic disease found in remote Andean areas. As an overlooked disease, knowledge of innate immune responses to Bartonella bacilliformis, the etiological agent, is scarce. This study aimed to evaluate the cytokine response to B. bacilliformis using in vitro human peripheral blood mononuclear cells (PBMCs) stimulations. PBMCs from naive adults were isolated by gradient centrifugation and cocultured with heat-inactivated (HI) B. bacilliformis at different incubation times (3, 6, 12, 24, and 36 h). Cytokines, chemokines, and growth factors were determined in culture supernatants by multiplex fluorescent bead-based quantitative suspension array technology. During the first 36 h, a proinflammatory response was observed, including tumor necrosis factor-α, interleukin (IL)-1α, IL-1ß, interferon-α2, and IL-6, followed by an anti-inflammatory response mainly related to IL-1RA. Moreover, high expression levels of chemokines IL-8, monocyte chemoattractant protein-1α, and macrophage inflammatory protein (MIP)-1ß were detected from 3 h poststimulation and MIP-1α was detected at 24 h. Some growth factors, mainly granulocyte macrophage colony-stimulating factor and granulocyte colony-stimulating factor, and in minor concentrations vascular endothelial growth factor, epidermal growth factor, and eotaxin, were also detected. Innate response to HI B. bacilliformis stimulation consists of a rapid and strong proinflammatory response characterized by a wide range of cytokines and chemokines followed by an anti-inflammatory response and increased specific growth factors.
Assuntos
Bartonella bacilliformis , Adulto , Humanos , Bartonella bacilliformis/fisiologia , Fator A de Crescimento do Endotélio Vascular , Leucócitos Mononucleares , Citocinas , Anti-InflamatóriosRESUMO
RESUMEN Objetivo. Evaluar in silico y a nivel serológico el potencial antigénico del dominio extracelular recombinante de la proteína de ensamblaje de lipopolisacáridos - D (LptD) de Bartonella bacilliformis (dexr_LptD). Materiales y métodos. Mediante el análisis in silico se realizó la selección de una proteína de B. bacilliformis con potencial antigénico e inmunogénico. El gen de la proteína seleccionada se clonó en Escherichia coli TOP10 y se expresó en Escherichia coli BL21 (DE3) pLysS. La proteína recombinante fue expresada usando isopropil-β-D-1-tiogalactopiranósido (IPTG) y se optimizaron las condiciones de inducción. Por último, se purificó con resina Ni-IDA (His60 Ni Superflow) y se realizó un ensayo de Western Blot. Resultados. In silico, la proteína seleccionada fue LptD por estar localizada en la membrana externa y ser antigénica e inmunogénica. Las condiciones optimizadas para la inducción del dexr_LptD fueron 0,5 mM IPTG, 16 h, medio TB (Terrific Broth), etanol al 3% (v/v), 28 ºC, OD600: 1-1,5 y 200 r.p.m. La purificación se realizó en condiciones denaturantes a pequeña escala y se obtuvo 2,6 µg/mL de dexr_LptD parcialmente purificada. El ensayo de Western Blot mostró una reacción positiva entre los sueros provenientes de pacientes con la enfermedad de Carrión y dexr_LptD, ello evidencia la antigenicidad del dexr_LptD. Conclusiones. El dexr_LptD muestra antigenicidad in silico y a nivel serológico, estos resultados son base para posteriores estudios sobre candidatos vacunales contra la enfermedad de Carrión.
ABSTRACT Objective. To evaluate in silico and at the serological level the antigenic potential of the recombinant extracellular domain of the lipopolysaccharide assembly protein - D (LptD) of Bartonella bacilliformis (dexr_LptD). Materials and Methods. Through in silico analysis, we selected a B. bacilliformis protein with antigenic and immunogenic potential. The selected protein gene was cloned into Escherichia coli TOP10 and expressed in Escherichia coli BL21 (DE3) pLysS. Recombinant protein was expressed using isopropyl-β-D-1-thiogalactopyranoside (IPTG) and induction conditions were optimized. Finally, it was purified with Ni-IDA resin (His60 Ni Superflow) and a Western Blot assay was conducted. Results. In silico, the selected protein was LptD because it is located in the outer membrane and is antigenic and immunogenic. Optimized conditions for dexr_LptD induction were 0.5 mM IPTG, 16 hours, TB (Terrific Broth) medium, 3% (v/v) ethanol, 28 ºC, OD600: 1-1.5 and 200 rpm. Purification was carried out under denaturating conditions on a small scale and we obtained 2.6 μg/mL of partially purified dexr_LptD. The Western Blot assay showed a positive reaction between the sera from patients with Carrión's Disease and dexr_LptD, which shows the antigenicity of dexr_LptD. Conclusions. The dexr_LptD shows antigenicity both in silico and at the serological level, these results are the basis for further studies on vaccine candidates against Carrion's Disease.
Assuntos
Proteínas Recombinantes , Clonagem de Organismos , Bartonella bacilliformis , Infecções por Bartonella , Biologia Computacional , Imunogenicidade da VacinaRESUMO
The presence of amino acid changes in GyrA, GyrB, ParC, ParE, and in a proposed chromosomal chloramphenicol acetyl transferase (CAT), as well as mutations at 23S rRNA, were established by PCR and sequencing in 38 B. bacilliformis clinical isolates from four different areas in Peru. Eighteen out of 24 (75%) isolates showing ciprofloxacin resistance for both disk-diffusion and e-test presented amino acid substitutions in GyrA (G89C, six isolates, A91V, 1 isolate) GyrB (S474F, 10 isolates) or both (GyrA D95N and GyrB S474F, one isolate). Two out of 14 susceptible isolates presented amino acid substitutions at GyrB (S474F) or a double substitution GyrA D95N and GyrB S474F. Of note, ciprofloxacin-resistant isolates were recovered in the four areas studied. No amino acid change was observed at ParC or ParE. Only one isolate showed chloramphenicol resistance, but no alteration was present in either 23S rRNA or CAT. B. bacilliformis resistant to quinolones are extended throughout Peru, with amino acid substitutions at GyrA or GyrB as the main, albeit not exclusive, cause. B. bacilliformis seems to have an apparent facility to develop mutations on GyrB outside the classical positions 91, 95 of GyrA and 85, 88 of ParC.
RESUMO
BACKGROUND Carrion's disease (CD) is a neglected biphasic illness caused by Bartonella bacilliformis, a Gram-negative bacteria found in the Andean valleys. The spread of resistant strains underlines the need for novel antimicrobials against B. bacilliformis and related bacterial pathogens. OBJECTIVE The main aim of this study was to integrate genomic-scale data to shortlist a set of proteins that could serve as attractive targets for new antimicrobial discovery to combat B. bacilliformis. METHODS We performed a multidimensional genomic scale analysis of potential and relevant targets which includes structural druggability, metabolic analysis and essentiality criteria to select proteins with attractive features for drug discovery. FINDINGS We shortlisted seventeen relevant proteins to develop new drugs against the causative agent of Carrion's disease. Particularly, the protein products of fabI, folA, aroA, trmFO, uppP and murE genes, meet an important number of desirable features that make them attractive targets for new drug development. This data compendium is freely available as a web server (http://target.sbg.qb.fcen.uba.ar/). MAIN CONCLUSION This work represents an effort to reduce the costs in the first phases of B. bacilliformis drug discovery.
Assuntos
Humanos , Infecções por Bartonella/tratamento farmacológico , Bartonella bacilliformis/efeitos dos fármacos , Antibacterianos/uso terapêutico , DNA Bacteriano/isolamento & purificação , DNA Bacteriano/genética , Reação em Cadeia da Polimerase , Genômica , Bartonella bacilliformis/isolamento & purificação , Bartonella bacilliformis/genéticaRESUMO
BACKGROUND: Carrion’s disease (CD) is a neglected biphasic illness caused by Bartonella bacilliformis, a Gram-negative bacteria found in the Andean valleys. The spread of resistant strains underlines the need for novel antimicrobials against B. bacilliformis and related bacterial pathogens. OBJECTIVE: The main aim of this study was to integrate genomic-scale data to shortlist a set of proteins that could serve as attractive targets for new antimicrobial discovery to combat B. bacilliformis. METHODS: We performed a multidimensional genomic scale analysis of potential and relevant targets which includes structural druggability, metabolic analysis and essentiality criteria to select proteins with attractive features for drug discovery. FINDINGS: We shortlisted seventeen relevant proteins to develop new drugs against the causative agent of Carrion’s disease. Particularly, the protein products of fabI, folA, aroA, trmFO, uppP and murE genes, meet an important number of desirable features that make them attractive targets for new drug development. This data compendium is freely available as a web server (http://target.sbg.qb.fcen.uba.ar/). MAIN CONCLUSION: This work represents an effort to reduce the costs in the first phases of B. bacilliformis drug discovery.
RESUMO
RESUMEN Objetivos. Diseñar y evaluar una proteína multiepítope como candidato a vacuna contra la enfermedad de Carrión. Materiales y métodos. Mediante herramientas bioinformáticas se seleccionó epítopes de proteínas de membrana externa y se diseñó una proteína multiepítope. El gen de la proteína multiepítope fue subclonado en el plásmido de expresión pET28b y transformado en E. coli BL21 pLys. La proteína multiepítope fue expresada usando isopropil-β-D-1-tiogalactopiranósido y purificada usando resina. Esta proteína purificada fue utilizada para inmunizar ratones BALB/c y se obtuvo anticuerpos policlonales. Se realizaron ensayos de invasión in vitro usando una cepa de Bartonella bacilliformis (B. bacilliformis) a eritrocitos humanos. Resultados. La proteína multiepítope M1 presenta epítopes conservados entre aislamientos de B. bacilliformis, no tóxicos, no homólogos a proteínas humanas y superficiales. Los ratones inmunizados presentaron niveles de anticuerpos IgG capaces de reducir in vitro la tasa de invasión de B. bacilliformis a eritrocitos humanos. Conclusiones. La proteína multiepítope M1 podría servir como candidato a vacuna contra la enfermedad de Carrión; sin embargo, se requiere de más estudios para caracterizar el uso de este antígeno como vacuna.
ABSTRACT Objectives. To design and assess a multiepitopic protein as a candidate for a vaccine against Carrion disease. Materials and Methods. Using bioinformatics tools, epitopes of external membrane proteins were selected and a multiepitopic protein was designed. The multiepitopic protein gene was subcloned into the expression plasmid pET28b and transformed into E. coli BL21 pLys. The multiepitopic protein was expressed using isopropyl-β-D-1-thiogalactopyranoside and purified using resin. This purified protein was used to immunize BALB/c mice obtaining polyclonal antibodies. In vitro invasion assays were conducted using a strain of Bartonella bacilliformis (B. bacilliformis) in human red blood cells. Results. The multiepitopic protein M1 presents preserved epitopes between isolates of B. bacilliformis with are non-toxic, and not homologous to human and surface proteins. Immunized mice presented IgG antibody levels capable of reducing in vitro the rate of invasion of B. bacilliformis into human red blood cells. Conclusions. Multiepitopic protein M1 may serve as a candidate for a Carrion disease vaccine; however, more studies are needed to characterize the use of this antigen as a vaccine.
Assuntos
Animais , Feminino , Proteínas de Bactérias/biossíntese , Infecções por Bartonella/prevenção & controle , Vacinas Bacterianas/biossíntese , Desenho de Fármacos , Biologia Computacional , Camundongos Endogâmicos BALB C , EpitoposRESUMO
Bartonella bacilliformis has recently been described in Amblyomma scalpturatum, Amblyomma ovale and Rhipicephalus microplus collected from wild animals in the Peruvian region of Madre de Dios. In this communication, I will discuss the results of a recent study by del Valle-Mendoza et al. together with the B. bacilliformis epidemiology. Following my argumentation, I consider the presence of this microorganism in the above ticks improbable.
Assuntos
Infecções por Bartonella/microbiologia , Bartonella bacilliformis/fisiologia , Carrapatos/microbiologia , Animais , Infecções por Bartonella/epidemiologia , Bartonella bacilliformis/genética , Bartonella bacilliformis/isolamento & purificação , Geografia , Humanos , Peru/epidemiologia , Carrapatos/classificaçãoRESUMO
BACKGROUND: Acute febrile illness (AFI) represent a significant health challenge in the Peruvian Amazon basin population due to their diverse etiologies and the unavailability of specific on-site diagnostic methods, resulting in underreporting of cases. In Peru, one of the most endemic regions to dengue and leptospirosis is Madre de Dios, a region also endemic to emergent bacterial etiologic agents of AFI, such as bartonellosis and rickettsiosis, whose prevalence is usually underreported. We aimed to molecularly identify the presence of Leptospira spp., Bartonella bacilliformis, and Rickettsia spp. by Polymerase Chain Reaction in serum samples from patients with AFI from Puerto Maldonado-Madre de Dios in Peru. METHODS: Serum samples from patients with acute febrile illness were analyzed by real-time PCR for detecting the presence of Bartonella bacilliformis, Leptospira spp. and Rickettsia spp. RESULTS: Bartonella bacilliformis was the most prevalent bacteria identified in 21.6% (30/139) of the samples, followed by Leptospira spp. in 11.5% (16/139) and Rickettsia spp. in 6.5% (9/139) of the samples. No co-infections were observed between these bacteria. The most frequent symptoms associated with fever among all groups, were headaches, myalgias, and arthralgias. We found no statistically significant differences in the clinical presentation between patients infected with each bacterium. CONCLUSIONS: In a previous study, we shown the presence of dengue, chikungunya, Zika and oropouche virus. We were able to identify these pathogens in 29.5% of all the samples, with chikungunya and OROV as the most frequently found in 9.4 and 8.6% of all the samples, respectively. In this study we show that B. bacilliformis (21.6%), Leptospira spp. (11.5%) and Rickettsia spp. (6.5%) accounted for the main etiologies of AFI in samples from Puerto Maldonado-Madre de Dios, Perú. Our analysis of their clinical presentation, further shows the importance of implementing more sensitive and specific on-site diagnostic tools in the national surveillance programs.This study confirms that the un-specificity of signs and symptoms is not only associated with arboviral infections, but also with the clinical presentation of endemic bacterial infections.
Assuntos
Infecções por Bartonella , Bartonella bacilliformis/genética , Leptospira/genética , Leptospirose , Infecções por Rickettsia , Rickettsia/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Infecções por Bartonella/diagnóstico , Infecções por Bartonella/epidemiologia , Infecções por Bartonella/genética , Infecções por Bartonella/microbiologia , Bartonella bacilliformis/isolamento & purificação , Criança , Pré-Escolar , Coinfecção , Estudos Transversais , Feminino , Febre/diagnóstico , Febre/epidemiologia , Febre/microbiologia , Humanos , Lactente , Recém-Nascido , Leptospira/isolamento & purificação , Leptospirose/diagnóstico , Leptospirose/epidemiologia , Leptospirose/microbiologia , Masculino , Pessoa de Meia-Idade , Tipagem Molecular , Peru/epidemiologia , Prevalência , Reação em Cadeia da Polimerase em Tempo Real , Rickettsia/isolamento & purificação , Infecções por Rickettsia/diagnóstico , Infecções por Rickettsia/epidemiologia , Infecções por Rickettsia/microbiologia , Rios , Sensibilidade e Especificidade , Adulto JovemRESUMO
OBJECTIVE: To study the presence of Bartonella bacilliformis in ticks collected from two wild mammals in Madre de Dios, Peru. RESULTS: A total of 110 ticks were collected. Among the 43 Amblyomma spp. extracted from the 3 Tapirus terrestris only 3 were positive for B. bacilliformis. In addition, 12 out of the 67 Rhipicephalus (Boophilus) microplus obtained from the 3 Pecari tajacu were positive for B. bacilliformis. For the first time B. bacilliformis have been detected in arthropods other than Lutzomyia spp. Further studies are required to elucidate the possible role of ticks in the spread of South American Bartonellosis.
Assuntos
Bartonella bacilliformis/isolamento & purificação , Carrapatos/microbiologia , Animais , Infecções por Bartonella , Bartonella bacilliformis/genética , Mamíferos , PeruRESUMO
All the studies published including Bartonella bacilliformis MLST data, as well as all B. bacilliformis genomes present in GenBank were analyzed. Overall 64 isolates and their geographical distribution were analyzed, and 14 different MLST patterns were observed. The results highlight the need for expanding the MLST studies and adding a higher number of isolates from all endemic areas.
Assuntos
Infecções por Bartonella/epidemiologia , Bartonella bacilliformis/genética , DNA Bacteriano/genética , Filogenia , Infecções por Bartonella/microbiologia , Bartonella bacilliformis/classificação , Bartonella bacilliformis/isolamento & purificação , Bases de Dados de Ácidos Nucleicos , Feminino , Genótipo , Humanos , Masculino , Tipagem de Sequências Multilocus , Peru/epidemiologiaRESUMO
OBJECTIVE: This study investigated an outbreak of Bartonellosis in a coastal region in Peru. RESULTS: A total of 70 (n = 70) samples with clinical criteria for the acute phase of Bartonellosis and a positive peripheral blood smear were included. 22.85% (n = 16) cases of the samples were positive for Bartonella bacilliformis by PCR and automatic sequencing. Of those positive samples, 62.5% (n = 10) cases were positive only for B. bacilliformis and 37.5% (n = 6) cases were positive to both Mycobacterium spp. and B. bacilliformis. The symptom frequencies were similar in patients diagnosed with Carrion's disease and those co-infected with Mycobacterium spp. The most common symptoms were headaches, followed by malaise and arthralgia.
Assuntos
Infecções por Bartonella/epidemiologia , Infecções por Bartonella/fisiopatologia , Bartonella bacilliformis/isolamento & purificação , Surtos de Doenças , Infecções por Mycobacterium/epidemiologia , Infecções por Mycobacterium/fisiopatologia , Mycobacterium/isolamento & purificação , Adolescente , Adulto , Idoso , Criança , Coinfecção , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Peru/epidemiologia , Adulto JovemRESUMO
Abstract Infective endocarditis due to Bartonella bacilliformis is rare. A 64-year-old woman, without previous heart disease, presented with 6 weeks of fever, myalgias, and arthralgias. A systolic murmur was heard on the tricuspid area upon examination, and an echocardiogram showed endocardial lesions in the right atrium. Bartonella bacilliformis was isolated in blood cultures, defining the diagnosis of infective endocarditis using Duke's criteria. Subsequently, the patient developed clinical and laboratory features compatible with antineutrophil cytoplasmic antibody-associated vasculitis. This case presents an uncommon complication of B. bacilliformis infection associated with the development of systemic vasculitis.
Assuntos
Humanos , Feminino , Infecções por Bartonella , Bartonella bacilliformis/isolamento & purificação , Endocardite Bacteriana/microbiologia , Ensaio de Imunoadsorção Enzimática , Ecocardiografia Transesofagiana , Endocardite Bacteriana/diagnóstico por imagem , Vasculite Sistêmica/microbiologia , Pessoa de Meia-IdadeRESUMO
The genus Bartonella contains >40 species, and an increasing number of these Bartonella species are being implicated in human disease. One such pathogen is Bartonella ancashensis, which was isolated in blood samples from 2 patients living in Caraz, Peru, during a clinical trial of treatment for bartonellosis. Three B. ancashensis strains were analyzed by using whole-genome restriction mapping and high-throughput pyrosequencing. Genome-wide comparative analysis of Bartonella species showed that B. ancashensis has features seen in modern and ancient lineages of Bartonella species and is more related to B. bacilliformis. The divergence between B. ancashensis and B. bacilliformis is much greater than what is seen between known Bartonella genetic lineages. In addition, B. ancashensis contains type IV secretion system proteins, which are not present in B. bacilliformis. Whole-genome analysis indicates that B. ancashensis might represent a distinct Bartonella lineage phylogenetically related to B. bacilliformis.
Assuntos
Infecções por Bartonella/microbiologia , Bartonella/genética , Genoma Bacteriano , Adolescente , Adulto , Bartonella/classificação , Infecções por Bartonella/epidemiologia , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Peru/epidemiologia , Filogenia , Adulto JovemRESUMO
Carrion's disease is a neglected tropical disease caused by Bartonella bacilliformis, a vector-borne pathogen restricted to the Andean valleys of Peru, Ecuador and Colombia. Carrion's disease is a biphasic illness; in the acute phase the case-fatality rate can be as high as 88 %, related to high parasitemia, arriving to almost all erythrocytes, and secondary bacterial infections close related with the development of transient immunosuppression in the earlier illness phases. In addition, there are an undefined number of asymptomatic carriers that are reservoirs of the etiological agent of Carrion's disease in endemic areas, they make take into account due to they are the perpetuators of this disease. The actual scenario of Carrion's disease, in which the illness is arriving to new areas, due to the expansion of the vector's distribution, suggests that now may be a crucial time to design a strategy focusing on its elimination.
Assuntos
Infecções por Bartonella/microbiologia , Infecções por Bartonella/prevenção & controle , Bartonella bacilliformis/fisiologia , Doenças Negligenciadas/microbiologia , Doenças Negligenciadas/prevenção & controle , Colômbia , Equador , Humanos , PeruRESUMO
La enfermedad de Carrión presenta clásicamente dos fases clínicas: la fiebre de la Oroya y la verruga peruana. La teoría unicista reconoce que ambas formas clínicas pertenecen a la misma enfermedad, la obtención de este conocimiento representó un duro desafío para la ciencia peruana y mundial. A través de un histórico experimento en 1885, Daniel Alcides Carrión obtuvo la evidencia clínica-experimental que sentó las bases de la teoría unicista; el científico diseñó esta teoría solo hasta las horas finales de su experimento, por lo cual su hallazgo es considerado un acto de serendipia. La demostración bacteriológica de la teoría la realizaría Hideyo Noguchi en 1926; Carrión brindó además evidencia de la transmisibilidad de esta infección y debido a este aporte científico se convirtió en el símbolo de la medicina peruana.
Carrion's disease typically presents two clinical phases: Oroya fever and Peruvian wart. Unicist theory recognizes that both clinical forms belong to the same disease. Obtaining this knowledge represented a challenge for both the Peruvian and the world science. In 1885, through a historical experiment, Daniel Alcides Carrión obtained the clinical-experimental evidence that work as the foundations of this theory. Carrion did not design this theory but only in the final hours of his experiment; therefore his finding is considered as an act of serendipity. Bacteriological demonstration of the theory was made in 1926 by Hideyo Noguchi. Carrion also provided evidence showing the transmission of this infection. Due to its scientific contribution Carrión became the symbol of the Peruvian medicine.
RESUMO
Daniel Carrion, a sixth-year medical student, died while investigating the effects of self-inoculation of the causative organism of Oroya Fever and Bartonellosis and thereby contributed to understanding of the disease before the organisms had been identified.
Assuntos
Autoexperimentação/história , Infecções por Bartonella/história , Bartonella bacilliformis/isolamento & purificação , Infecções por Bartonella/microbiologia , História do Século XIX , Humanos , Masculino , Peru , Estudantes de Medicina/históriaRESUMO
Se reporta el caso de un adulto varón de 56 años, procedente de Sihuas (Ancash) que acude al Hospital Nacional Dos de Mayo con cuadro clínico caracterizado por un síndrome ictérico febril anemizante, en cuya evolución tórpida de solo tres días de hospitalizado presenta anemia severa (Hb: 11,8 a 6,7 g/dL) así como mialgias generalizadas asociadas a un empeoramiento brusco de la ictericia correlacionada con elevación de bilirrubinas (BT: 3,2 a 19,6 mg/dL a predominio directo) y deterioro general del paciente; por ello, se realizó la investigación microbiológica donde se encontró frotis y cultivo positivo para Bartonella bacilliformis, posteriormente, la serología (microaglutinación) resultó positiva para Lesptospira serovar Pomona. El paciente recibe tratamiento antibiótico específico (ceftriaxona y ciprofloxacino), se le transfunde dos paquetes globulares, mejora clínicamente y es dado de alta.
The case of an adult, 56 year old male is reported, from Sihuas (Ancash) who presented at Hospital Nacional Dos de Mayo with fever, jaundice and anemia. In three days of hospitalization, he developed severe anemia (Hb: 11.8 to 6.7 g / dL) and generalized myalgias associated with a sudden worsening jaundice correlated with elevated bilirubin (TB 3.2 to 19.6 mg / dL direct dominance) and general deterioration. Microbiological smear and culture were positive for Bartonella bacilliformis. Subsequent serology (microagglutination) was positive for Lesptospira serovar Pomona. The patient was given specific antibiotic therapy (ceftriaxone and ciprofloxacin), transfused two units of packed red blood cells, improved clinically and was discharged.
Assuntos
Humanos , Masculino , Pessoa de Meia-Idade , Infecções por Bartonella/complicações , Leptospirose/complicações , Doença Aguda , Índice de Gravidade de DoençaRESUMO
Bartonella bacilliformis es el agente etiológico de la Enfermedad de Carrión, endémica del Perú. Pocas investigaciones han sido realizadas acerca de los genes asociados a la resistencia antimicrobiana en aislados clínicos de este patógeno. Estos genes no están caracterizados molecularmente, ni se conoce la región asociada a dicha resistencia. Por ello, el objetivo del este trabajo fue caracterizar molecularmente la región determinante de la resistencia a las quinolonas (QRDR) en la topoisomerasa IV, que está codificada por los genes parC y parE, así como también desarrollar una prueba de susceptibilidad antimicrobiana para B. bacilliformis. Las muestras sanguíneas de 65 pacientes procedentes de La Libertad, Cusco, Ancash y Piura, se sembraron en placas de agar sangre e incubaron a 30 °C con 5% CO2. Luego se procedió a: (1) determinar la susceptibilidad antimicrobiana y (2) extraer el DNA genómico, amplificar los genes mencionados, secuenciarlos y analizarlos mediante herramientas bioinformáticas. Se obtuvieron 6 cultivos positivos. Los aislados fueron sensibles a la ciprofloxacina (excepto uno procedente de Quillabamba-Cusco, que presentó susceptibilidad disminuida) y resistentes al ácido nalidíxico. Del análisis de las secuencias aminoacídicas de ParC y ParE de B. bacilliformis se concluye que presentan diferencias aminoacídicas en comparación con las secuencias de las proteínas respectivas de E. coli K12 MG1655, que probablemente confieran resistencia al ácido nalidíxico pero no a la ciprofloxacina. Se determinó que las QRDR de las proteínas ParC y ParE de B. bacilliformis están comprendidas entre los aminoácidos 67 al 118 y 473 al 530, respectivamente. El antibiograma y la concentración mínima inhibitoria se evalúan mejor usando inóculos a escala 1 de McFarland y a los 6 días de incubación
Bartonella bacilliformis is the etiologic agent of Carrion's disease, which if endemic to Peru. Studies on antimicrobial resistance genes from clinical isolates of this pathogen are scarce, and the molecular characteristics of these genes and their region resistance-associated are currently unknown. In this work we made the molecular characterization of the quinolone-resistance, and establish the region (QRDR) for the topoisomerase IV, which is encoded by the parC and parE genes, as well as develop an antimicrobial susceptibility test for B. bacilliformis. 65 Blood samples from La Libertad, Cusco, Ancash and Piura were processed on Blood Agar plates and incubated at 30 °C, 5% CO2. The antimicrobial susceptibility was determined, then the genomic DNA extracted, aforementioned genes amplified, their sequence determined and it analyzed using bioinformatics tools. Six positive cultures were obtained. The isolates were susceptible to Ciprofloxacin (except one strain from Quillabamba _ Cusco, which showed decreased susceptibility) and were resistant to Nalidixic Acid. From the sequence analysis of B. bacilliformis ParC and ParE there have been shown amino acid differences compared to the respective protein sequences from E. coli K12 MG1655, which is likely to confer resistance to Nalidixic Acid but not to Ciprofloxacin. It was determined that B. bacilliformis ParC and ParE proteins QRDRs are comprised between amino acids 67 to 118 and 473 to 530, respectively. The antibiogram and the minimal inhibitory concentration are best assessed using the #1 McFarland standards after a 6-day incubation period
RESUMO
OBJETIVO: Evaluar el conocimiento, las prácticas y las actitudes sobre la enfermedad de Carrión (bartonelosis) en poblaciones rurales con antecedentes de brotes epidémicos. MÉTODOS: Estudio transversal realizado en las poblaciones de Huaripampa, Orcosh y Opayaco (Ancash, Perú) durante el mes de julio del 2010, utilizando una encuesta para evaluar el conocimiento, las prácticas y las actitudes sobre la enfermedad de Carrión. Se entrevistó solo a mayores de 18 años que tuviesen una antigüedad mínima de un año residiendo en esos poblados. RESULTADOS: Se realizaron encuestas a un total de 276 pobladores, de los cuales 36,6% fueron hombres y 72,5% no había oído hablar de la enfermedad de Carrión. Entre los que la conocían, la mayoría (38,7%) indicó que se contagia mediante picaduras, mientras que 26% manifestó no conocer la forma de contagio. Con respecto a las acciones que deberían tomarse en caso de presentar la enfermedad de Carrión, 69,3% de los encuestados refirieron que acudirían a un centro de salud y más de la mitad informó haber recomendado prácticas preventivas a familiares o conocidos en el pasado. CONCLUSIONES: A pesar de tratarse de comunidades históricamente afectadas por la enfermedad de Carrión, los pobladores de estas tres comunidades no mostraron un nivel adecuado de conocimientos, actitudes o prácticas para su prevención. En vista de ello, y teniendo en cuenta que este padecimiento ha presentado brotes cíclicos, se hace perentorio diseñar e implementar un programa que eduque a los habitantes de estos poblados -poniendo el foco en el papel de los usos y costumbres- sobre cómo prevenir la enfermedad de Carrión, así como el resto de las principales enfermedades endémicas de la región.
OBJECTIVE: To assess knowledge, attitudes, and practices about Carrión's disease (bartonellosis) in rural communities with a history of epidemic outbreaks. METHODS: A cross-sectional survey study was conducted in the communities of Huaripampa, Orcosh, and Opayaco (Ancash, Peru) in July 2010, to assess knowledge, attitudes, and practices about Carrión's disease. Interviewees were aged >18 years and had resided in the community for at least one year. RESULTS: A total of 276 residents were surveyed; 36.6% were men, and 72.5% had not heard of Carrión's disease. Of those familiar with it, most (38.7%) said that it spreads through bites, and 26% did not know how it spreads. Regarding actions to take if Carrión's disease develops, 69.3% of respondents said they would go to a health center and over half reported having recommended preventive practices to relatives or acquaintances in the past. CONCLUSIONS: Even though these three communities have a history of Carrión's disease, their inhabitants did not demonstrate adequate knowledge, attitudes, or practices for preventing it. In view of this, and given that this illness has cyclical outbreaks, it is imperative to design and implement a program to educate residents of these communities-focusing on the role of habits and customs-about how to prevent Carrión's disease, as well as the region's other main endemic diseases.
Assuntos
Humanos , Masculino , Feminino , Adolescente , Adulto , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Adulto Jovem , Infecções por Bartonella , Conhecimentos, Atitudes e Prática em Saúde , Infecções por Bartonella/epidemiologia , Estudos Transversais , Surtos de Doenças , Peru/epidemiologia , Inquéritos e Questionários , Saúde da População RuralRESUMO
Objetivos: Describir el tratamiento para infección por Bartonella bacilliformis, en Caraz, Ancash. Diseño: Estudio observacional, transversal y retrospectivo. Lugar: Caraz, Ancash, Perú. Pacientes: Quinientos dieciocho pacientes. Intervenciones: Recolección de datos, entre enero 2004 y marzo 2005, por medio de historias clínicas realizadas en el Hospital de Caraz. Principales medidas de resultados: Esquemas de tratamiento para infección por Bartonella bacilliformis. Resultados: Se registró 248 pacientes en fase aguda. En 215 (86,7 por ciento), se indicó cloramfenicol para su tratamiento y en todos ellos no se registró la dosis de carga de 50 mg/kg, por 3 días; 164 (66,1 por ciento) pacientes necesitaron una dosis de cloramfenicol superior a 25 mg/kg. En fase eruptiva, se registró 270 pacientes; en 260 (96,3 por ciento) se indicó rifampicina y 222 (82,2 por ciento) sobrepasaron los 21 días de duración de tratamiento con dicho antibiótico. Cloramfenicol obtuvoun 89 por ciento de eficacia en curación clínica y rifampicina, 93,1 por ciento. Conclusiones: En la población estudiada, existió una marcada tendencia a aumentar la dosis de cloramfenicol e incrementar el número de días de tratamiento con rifampicina, en fase aguda y eruptiva, respectivamente, de la Enfermedad de Carrión.
Objectives: To describe Bartonella bacilliformis' infection treatment, in Caraz, Ancash. Design: Observational, transverse and retrospective study. Setting: Caraz, Ancash, Peru. Patients: Five hundred and eighteen patients. Interventions: Study of clinical histories data from January 2004 through March 2005 at Caraz Hospital. Main outcome measures: Treatment outlines for Bartonella bacilliformis infection. Results: In the acute phase, 248 patients were registered; in 215 (86,7 per cent) chloramphenicol treatment was indicated but the 50 mg/kg loading dose by 3 days was not prescribed; 164 (66,1 per cent) patients needed a higher chloramphenicol dose of 25 mg/kg. In the eruptive phase, 270 patients were registered; in 260 (96,3 per cent) rifampicin was indicated and 222 (82,2 per cent) for more than the 21 treatment days suggested for this antibiotic. Clinical cure was obtained with chloramphenicol in 89 per cent and 93,1 per cent with rifampicin. Conclusions: In the population studied there was a tendency to increase both chloramphenicol dose and rifampicin number of treatment days respectively in the acute and eruptive phases of Carrion's disease.