The influence of growth time on the identification of Bartonella henselae strains by MALDI-TOF mass spectrometry

ABSTRACT Bartonella spp. are bacteria responsible for neglected diseases worldwide. Bartonella henselae is the species most associated with human infections. It is associated with a large spectrum of clinical manifestations and is potentially fatal. The identification of Bartonella spp. is considered a challenge in clinical routine. These bacteria are fastidious, and the time required to isolate them varies from one to six weeks. MALDI-TOF mass spectrometry has emerged as an application for research on Bartonella spp. , and has still been little explored. We investigated whether three different B. henselae strains with different growth times—14 and 28 days—could be correctly identified by MALDI-TOF mass spectra fingerprint comparison and matching. We found that the spectra from strains with different growth times do not match each other, leading to misidentification. We suggest creating database entries with multiple spectra from strains with different growth times to increase the chances of accurate identification of Bartonella spp. by MALD-TOF MS.

Encefalomielite aguda disseminada com provável etiologia de Bartonella henselae / Acute disseminated encephalomyelitis with probable Bartonella henselae etiology

RESUMO A encefalomielite aguda disseminada é uma doença rara, aguda, inflamatória e desmielinizante do sistema nervoso central, presumivelmente associada, em mais de três quartos dos casos, a infecções (virais, bacterianas ou inespecíficas) e imunizações ou sem qualquer antecedente indentificável. Habitualmente, apresenta um curso monofásico com início de sintomas inespecíficos na fase prodrómica, podendo evoluir com alterações neurológicas multifocais e até à perda total da acuidade visual. Descrevemos o caso de um menino de 9 anos com quadro inicial de edema de papila causado por encefalomielite aguda disseminada devido a Bartonella henselae. Apesar da gravidade da doença, o diagnóstico e o tratamento precoce proporcionaram bons desfechos.

ABSTRACT Acute disseminated encephalomyelitis is a rare, acute, inflammatory, and demyelinating disease of the central nervous system. Presumably associated in more than three quarters of cases by infections (viral, bacterial, or nonspecific) and immunizations or without any identifiable antecedent. It usually presents a monophasic course with onset of nonspecific symptoms in the prodromal phase and may evolve with multifocal neurological changes and even visual acuity loss. We describe a case of a 9-year-old boy with an initial picture of papillary edema caused by acute disseminated encephalomyelitis due to Bartonella henselae. Despite the severity of the disease, early diagnosis and treatment provided good outcomes.

Presumed acute posterior multifocal placoid pigmentary epitheliopathy associated with Bartonella infection / Epiteliopatia pigmentar placoide multifocal posterior aguda presumível associada com infecção por Bartonella

ABSTRACT To report a unique case of acute posterior multifocal placoid pigment epitheliopathy (APMPPE) in a patient with positive serology for Bartonella, presenting with ocular signs and symptoms not attributable to other diseases. A 27-year-old woman presented with decreased visual acuity in both eyes. Multimodal fundus image analysis was performed. A color fundus photograph of both eyes revealed peripapillary and macular yellow-white placoid lesions. The fundus autofluorescence of both eyes demonstrated hypo- and hyperautofluorescence of the macular lesions. Fluorescein angiography showed early-stage hypofluorescence and late staining of placoid lesions in both eyes. Spectral domain optical coherence tomography (SD-OCT) of both eyes revealed irregular elevations in the retinal pigment epithelium with the disruption of the ellipsoid zone on the topography of macular lesions. At 3 months after the treatment initiation for Bartonella infection, the placoid lesions became atrophic and hyperpigmented, and SD-OCT revealed loss of both the outer retinal layers and retinal pigment epithelium on the topography of macular lesions in both eyes.

RESUMO Caso de epiteliopatia pigmentada placoide multifocal posterior aguda presumida em paciente com sorologia positiva para Bartonella. Paciente feminina de 27 anos apresentou diminuição da acuidade visual em ambos os olhos. Análise multimodal de imagem foi realizada. A retinografia mostrou revelou lesões placoides amarelo-esbranquiçadas nas áreas peripapilar e macular de ambos os olhos. A autofluorescência demonstrou hipo e hiperautofluorescência em ambos os olhos, na mesma topografia das lesões detectadas na retinografia. A angiofluoresceínografia mostrou hipofluorescência na fase inicial do exame e hiperfluorescência tardia das lesões placoides em ambos os olhos. A tomografia de coerência óptica de domínio espectral de ambos os olhos revelou elevações irregulares do epitélio pigmentado da retina com descontinuação da zona elipsoide na área macular. Três meses após o início do tratamento para infecção por Bartonella, as lesões placoides tornaram-se atróficas e hiperpigmentadas, e a tomografia de coerência óptica revelou perda das camadas externas da retina e do epitélio pigmentado da retina na topografia das lesões maculares em ambos os olhos.

Variantes en la presentación clínica de la enfermedad por arañazo de gato en niños del hospital Garrahan / Variants in the clinical presentation of cat scratch disease in children at Garrahan Hospital

La infección por Bartonella henselae (BH) adopta diversas formas de presentación clínica en pediatría. Según la bibliografía la forma de presentación más frecuente en pacientes inmunocompetentes es la linfadenopatía única asociada a fiebre. En el 85 % de los casos se compromete un solo ganglio siendo los axilares y los epitrocleares los más frecuentemente involucrados. Existen otras formas de presentación menos frecuentes que debemos tener en consideración, para poder realizar un diagnóstico precoz e indicar un tratamiento adecuado si así lo requiere. El diagnóstico requiere de la sospecha clínica del equipo de salud tratante, junto al antecedente epidemiológico, los hallazgos clínicos del examen físico y la realización de serologías que incluyan el dosaje de inmunoglobulina M y G. Los objetivos del presente trabajo fueron reconocer las manifestaciones clínicas típicas y atípicas de la EAG por Bartonella henselae, describir la epidemiología, características clínicas y evolución de esta enfermedad que se presentaron en nuestro hospital. Se estudiaron un total de 187 pacientes. La media de edad fue de 7.6 años (rango 1-14); siendo 53.5% de género masculino. Las formas de presentación más frecuentes en nuestro trabajo fueron la adenitis y la fiebre. La mayoría recibió diversos esquemas de tratamiento antibiótico, secundario al retraso en el diagnóstico. La tasa de hospitalización fue muy baja, remitió con tratamiento ambulatorio con antibióticos o sin ellos (AU)

Bartonella henselae infection has different clinical presentations in pediatrics. According to the literature, the most common form of presentation in immunocompetent patients is single lymphadenopathy associated with fever. In 85 % of the cases a single lymph node is involved, with the axillary and epitrochlear nodes being the most commonly involved. There are other, less frequent, forms of presentation that should be taken into consideration in order to make an early diagnosis and indicate appropriate treatment if required. Diagnosis relies on clinical suspicion by the treating healthcare team, together with the epidemiological history, clinical findings on physical examination, and serology including immunoglobulin M and G dosage. The objectives of this study were to identify both the typical and atypical clinical manifestations of Bartonella henselae cat scratch disease, to describe the epidemiology, clinical characteristics, and outcomes of cases presenting at our hospital. A total of 187 patients were studied. The mean age was 7.6 years (range 1-14); 53.5% were male. The most frequent forms of presentation in our study were adenitis and fever. Most of them received different antibiotic treatment regimens due to delayed diagnosis. The hospitalization rate was very low and the disease typically resolved with outpatient treatment, with or without antibiotics (AU)

Detection of Bartonella henselae DNA in the blood of patients with livedoid vasculopathy

Abstract Background: Livedoid vasculopathy (LV) manifests as ulcers and atrophic white scars on the lower extremities. The main known etiopathogenesis is hypercoagulability with thrombus formation, followed by inflammation. Thrombophilia, collagen and myeloproliferative diseases may induce LV, but the idiopathic (primary) form predominates. Bartonella spp. may cause intra-endothelial infection and skin manifestations caused by these bacteria may be diverse, including leukocytoclastic vasculitis and ulcers. Objective: The aim of this study was to investigate the presence of bacteremia by Bartonella spp. in patients with difficult-to-control chronic ulcers diagnosed as primary LV. Methods: Questionnaires and molecular tests (conventional PCR, nested PCR and real-time PCR) were applied and liquid and solid cultures were performed in the blood samples and blood clot of 16 LV patients and 32 healthy volunteers. Results: Bartonella henselae DNA was detected in 25% of LV patients and in 12.5% of control subjects but failed to reach statistically significant differences (p = 0.413). Study limitations: Due to the rarity of primary LV, the number of patients studied was small and there was greater exposure of the control group to risk factors for Bartonella spp. infection. Conclusion: Although there was no statistically significant difference between the groups, the DNA of B. henselae was detected in one of every four patients, which reinforces the need to investigate Bartonella spp. in patients with primary LV.

Los aportes científicos extranjeros en el estudio de la Enfermedad de Carrión

El actual estado del conocimiento de la Enfermedad de Carrión, patología existente fundamentalmente en algunas zonas de la serranía peruana, se debe, en gran medida, a las investigaciones de los médicos de esa nacionalidad, sin embargo, galenos, otros científicos, viajeros, publicaciones y adelantos tecnológicos provenientes de otros países también han brindado aportes sobre esta entidad nosológica, algunos trascendentes y otros contraproducentes. El propósito de este artículo es reconocer tales contribuciones en el contexto de la evolución del conocimiento científico e histórico de dicha enfermedad.

The current state of knowledge of Carrión's Disease, a pathology that exists mainly in some areas of the Peruvian highlands, is due, to a large extent, to the investigations of doctors of that nationality, however, doctors, other scientists, travelers, publications, and technological advances from other countries have also provided contributions on this nosologically entity, some transcendent and others counterproductive. The purpose of this article is to acknowledge such contributions in the context of the evolution of scientific and historical knowledge of this disease.

Angiomatosis bacilar y micobacteriosis atípica en paciente con diagnóstico reciente de infección por el virus de la inmunodeficiencia humana / Bacillary Angiomatosis and Atypical Mycobacteriosis in a patient recently diagnosed with Human Immunodeficiency Virus infection

Múltiples infecciones oportunistas pueden manifestarse simultáneamente cuando la inmunosupresión es grave en pacientes infectados por el virus de la inmunodeficiencia humana. Se presenta un caso de coinfección de VIH, Bartonella spp y Mycobacterium kansasii, siendo escasos los reportes a la fecha de dicha asociación y aun más en pacientes que desconocen su condición de VIH

Multiple opportunistic infections can manifest simultaneously when immunosuppression is severe in patients infected with the human immunodeficiency virus. A case of co-infection with HIV, Bartonella spp and Mycobacterium kansasii is presented, with few reports to date of this association and even more so in patients who are unaware of their HIV status.

Enfermedad por arañazo de gato: experiencia en unhospital pediátrico de tercer nivel / Cat-scratch disease: Experience in a tertiary care children's hospital

Introducción. Bartonella henselae es el agente etiológico de la enfermedad por arañazo de gato. Afecta a niños y a adultos jóvenes. El espectro clínico es amplio; la forma de presentación más frecuente es la linfadenopatía única. El objetivo de este estudio fue analizar epidemiología, características clínicas y evolución de esta enfermedad en un hospital de alta complejidad de Argentina. Población y métodos. Estudio retrospectivo, descriptivo y observacional realizado en un hospital pediátrico de tercer nivel, desde el 01 de enero de 2019 hasta el 30 de junio de 2021. Se incluyeron niños de 0 a 16 años con clínica compatible y serología positiva. Resultados. Se incluyeron 150 niños, con una media de edad de 7,9 años ± 3,68. El 68,7 % refirió tener contacto con gatos. El motivo de consulta más frecuente fueron las adenopatías únicas (84,7 %) localizadas en cabeza y cuello. El síndrome febril sin foco motivó la consulta en el 15,5 % de los casos, con ecografía abdominal patológica en el 85,7 %. Presentó IgM e IgG positivas el 88 %. Con el resultado de la serología positiva, el 44 % recibió tratamiento antibiótico. Las adenopatías prolongadas fueron la principal causa de su instauración; el más utilizado fue la azitromicina (42,4 %). El 14 % (n = 21) requirió internación. Conclusiones. El diagnóstico implica sospecha clínica, nexo epidemiológico y exámenes complementarios. Su forma típica son las adenomegalias únicas localizadas en cabeza y cuello. Debido a la alta frecuencia de compromiso hepatoesplénico, la realización de ecografía abdominal estaría indicada en niños con fiebre.

Introduction. Bartonella henselae is the etiologic agent in cat-scratch disease. It affects children and young adults. The clinical spectrum is wide; the most common clinical presentation is a solitary lymphadenopathy. The objective of this study was to analyze the epidemiology, clinical features, and course of this disease in a tertiary care hospital in Argentina. Population and methods. Retrospective, descriptive, and observational study conducted at a tertiary care pediatric hospital from January 1st, 2019 to June 30 th, 2021. Children aged 0 to 16 years with compatible clinical signs and symptoms and positive serology were included. Results. A total of 150 patients were included; their mean age was 7.9 years ± 3.68. Of them, 68.7% reported having contact with cats. The most common reason for consultation was the presence of solitary lymphadenopathies (84.7%) in the head and neck. Febrile syndrome without source was the reason for consultation in 15.5% of cases, with a pathological abdominal ultrasound scan in 85.7%. IgM and IgG were positive in 88%. With the result of a positive serology test, 44% received antibiotic treatment. Protracted lymphadenopathy was the main reason for antibiotic treatment; the agent most commonly used was azithromycin (42.4%). Fourteen percent (n = 21) required hospitalization. Conclusions. Diagnosis is based on clinical suspicion, epidemiological history, and complementary testing. Its typical presentation is a solitary enlarged lymph node in the head and neck. Due to the high frequency of hepatosplenic involvement, an abdominal ultrasound scan would be indicated in children with fever.

Bartonelose: doença de importância para a saúde pública envolvendo a tríade homem-ambiente-animal / Bartonellosis: disease of importance for public health involving the man-environment-animal triad / Bartonelosis: enfermedad de importancia para la salud pública que involucra la triada hombre-ambiente-animal

A relação hospedeiro-parasita é caracterizada como uma interação alelobiótica construída por meio de processos evolutivo-adaptativos com hospedeiros assintomáticos. No ambiente silvestre é notório o equilíbrio desta relação, porém quando há intervenção antropogênica um ciclo enzoótico pode se estabelecer proporcionando o surgimento de enfermidades emergentes ou reemergentes. Dentre estes agentes etiológicos, a Bartonella spp. é um bacilo gram-negativo da classe Proteobacteria que apresentam tropismo por eritrócitos e células endoteliais, com infecção já descrita em animais das Ordens: Rodentia, Lagomorpha, Carnivora, Artiodactyla, Eulipotyphla e Chiroptera. A infecção pela bactéria pode estar associada à linfadenite, endocardite, angiomatose bacilar e peliose hepática em humanos. Treze espécies de Bartonella spp. são tidas como zoonóticas. O objetivo desta revisão está em apontar para a comunidade científica a bartonelose como uma doença de notificação obrigatória, assim como, os possíveis hospedeiros em animais domésticos e silvestres e sua etiopatogenia.(AU)

The host-parasite relationship is characterized as an allelobiotic interaction built through evolutionary-adaptive processes with asymptomatic hosts. In the wild environment, the balance of this relationship is notorious, but when there is anthropogenic intervention, an enzootic cycle can be established, providing the emergence of emerging or reemerging diseases. Among these etiologic agents, Bartonella spp. is a gram-negative bacillus of the Proteobacteria class that presents tropism for erythrocytes and endothelial cells, with infection already described in animals of the Orders: Rodentia, Lagomorpha, Carnivora, Artiodactyla, Eulipotyphla and Chiroptera. Infection by the bacterium may be associated with lymphadenitis, endocarditis, bacillary angiomatosis and peliosis hepatica in humans. Thirteen species of Bartonella spp. are considered zoonotic. The objective of this review is to point out to the scientific community bartonellosis as a notifiable disease, as well as the possible hosts in domestic and wild animals and their etiopathogenesis.(AU)

La relación hospedador-parásito se caracteriza por ser una interacción alelobiótica construida mediante procesos evolutivo-adaptativos con hospedadores asintomáticos. En el medio silvestre, el equilibrio de esta relación es notorio, pero cuando hay intervención antropogénica, puede establecerse un ciclo enzoótico, propiciando la aparición de enfermedades emergentes o reemergentes. Entre estos agentes etiológicos, Bartonella spp. es un bacilo gramnegativo de la clase Proteobacteria que presenta tropismo por eritrocitos y células endoteliales, con infección ya descrita en animales de los Órdenes: Rodentia, Lagomorpha, Carnivora, Artiodactyla, Eulipotyphla y Chiroptera. La infección por la bacteria puede estar asociada a linfadenitis, endocarditis, angiomatosis bacilar y peliosis hepática en humanos. Trece especies de Bartonella spp. se consideran zoonóticas. El objetivo de esta revisión es señalar a la comunidad científica la bartonelosis como enfermedad de declaración obligatoria, así como los posibles hospedadores en animales domésticos y salvajes y su etiopatogenia.(AU)

Infección por bartonella henselae en pacientes pediátricos. Lesión pseudotumoral de calota. Reporte de caso

Los tumores de calota en pacientes pediátricos poseen múltiples etiologías. Dentro de las causas pseudotumorales, las infecciones juegan un rol importante, siendo la osteomielitis por Bartonella henselae (Enfermedad por Arañazo de Gato) una posibilidad diagnóstica rara, pero que debe ser estudiada y descartada. Se presenta el caso de una lactante de 1 año, con lesión expansiva de calota, a nivel frontal derecho, hipervascularizada e infiltrativa. Se realizó estudio con ultrasonido, tomografía cerebral y cintigrama óseo. Se realizó resección quirúrgica completa de la lesión, con preservación de la duramadre y zona fontanelar, además de un cuidadoso trato con el seno sagital superior. Evolucionó sin complicaciones perioperatorias. El resultado de la biopsia fue compatible con proceso inflamatorio crónico, osteomielitis supurada. Tinción de Warthin Starry positiva sugerente de Bartonella henselae. Se descartó etiología tuberculosa y fúngica. Serología positiva para Bartonella henselae. La paciente completó antibioticoterapia, azitromicina y cotrimoxazol, con evolución clínica favorable.

Calvarial tumors in pediatric patients have multiple etiologies. Among the pseudotumoral causes, infections play an important role, being Bartonella henselae osteomyelitis (Cat Scratch Disease) a rare diagnostic possibility, but it should be studied and ruled out. We present the case of a 1 year old infant, with an expansive lesion of the calvaria, at right frontal level, hypervascularized and infiltrative. Ultrasound, brain tomography and bone scintigram were performed. Complete surgical resection of the lesion was performed, with preservation of the dura mater and fontanel area, in addition to a careful treatment with the superior sagittal sinus. The patient evolved without perioperative complications. The biopsy result was compatible with a chronic inflammatory process, suppurative osteomyelitis. Positive Warthin Starry stain suggestive of Bartonella henselae. Tuberculous and fungal etiology was ruled out. Positive serology for Bartonella henselae. The patient completed antibiotic therapy, azithromycin and cotrimoxazole, with favorable clinical evolution.

Bartonella henselae as a putative trigger for chronic type 2 leprosy reactions

Abstract Leprosy reactions are an acute inflammatory phenomenon that can arise before diagnosis, during treatment, or after cure of leprosy. These reactions are considered one of the main diseases that cause physical disabilities. Immunosuppressive treatment for these immune responses makes these patients susceptible to coinfections, which can trigger new leprosy reactions. The main objective of this study was to evaluate the occurrence of infection by Bartonella sp. in blood samples from 47 patients who had untreatable episodes of type 2 leprosy reactions for more than six months, comparing them with a control group. Cultures and molecular methods (PCR) were used. Amplicons from species-specific reactions and sequencing showed a higher prevalence of Bartonella henselae infection in patients, 19/47 (40.4 %), compared to control, 9/50 (18.0 %), p= 0.0149. Five patients accepted treatment for coinfection, and all showed improvement in leprosy reactions with treatment for B. henselae infection. We conclude that these bacteria can trigger chronic reactions of type 2 leprosy and should be investigated in these patients. Summary line Patients who have chronic type 2 leprosy reactions are more susceptible to Bartonella henselae infection than controls: 19/47 (40.4 %) compared 9/50 (18.0 %), p= 0.0149.

Enfermedad por arañazo de gato atípica: compromiso ocular y meníngeo por Bartonella henselae / Atypical cat scratch disease: Bartonella henselae ocular and meningeal involvement

Bartonella henselae es el agente etiológico de la enfermedad por arañazo de gato. Típicamente, se presenta como una linfadenopatía regional autolimitada y, con menor frecuencia, con compromiso sistémico y manifestaciones extraganglionares: hígado, bazo, hueso y ojo, entre otros. Se presenta un caso de enfermedad por arañazo de gato atípica en un paciente pediátrico inmunocompetente, en la que se evidenció compromiso meníngeo y ocular, este último como neurorretinitis. Se destaca la importancia de la búsqueda activa de complicaciones oculares en pacientes con compromiso sistémico por Bartonella henselae, que implica un cambio en el tratamiento y pronóstico de la enfermedad

Bartonella henselae is the etiologic agent of cat scratch disease. It typically presents as a self-limited regional lymphadenopathy and less frequently with systemic involvement and extranodal manifestations: liver, spleen, bone, eye, among others. A case of atypical cat scratch disease is presented in an immunocompetent pediatric patient, in which meningeal and ocular involvement was evidenced, the latter manifested as neuroretinitis. The importance of the active search for ocular complications in patients with systemic involvement by Bartonella henselae is highlighted, implying a change in the treatment and prognosis of the disease

Miositis por Bartonella henselae: caso clínico en una adolescente / Bartonella henselae myositis: case report in an adolescent

La miositis de origen vírico o bacteriano es frecuente en la edad pediátrica. Causa dolor muscular y debilidad, con fiebre y malestar general. Una causa es la infección por Bartonella henselae, bacteria implicada en la enfermedad por arañazo de gato que, a veces, causa afectación multisistémica. Se presenta el caso de una adolescente que acudió al servicio de urgencias por mialgia intensa, malestar, adelgazamiento y esplenomegalia. En el labortorio se observaron parámetros inflamatorios elevados. Refería contacto con un gato. Entre los estudios realizados, la resonancia magnética (RM) de miembros inferiores mostró una imagen compatible con miositis inflamatoria bilateral. En la RM abdominal, se observaron tres lesiones esplénicas no detectadas previamente y el fondo de ojo mostraba una lesión compatible con oclusión arterial retiniana o vasculitis. Se indicó tratamiento antibiótico por vía intravenosa durante 21 días con cefotaxima y cloxacilina, tras los cuales desaparecieron los signos y síntomas, aunque los reactantes inflamatorios persistieron elevados. Con base en el cuadro clínico (miositis + coriorretinitis + absceso esplénico) se pensó en una posible infección por B. henselae y se inició tratamiento oral con azitromicina y rifampicina durante 14 días. Luego del tratamiento, los valores de laboratorio fueron normales, así como la RM de control, y se constató una IgG positiva para la bacteria

Infectious myositis, whether viral or bacterial, is frequent in pediatric age. It causes muscle pain and weakness, associated with fever and general malaise. One cause is Bartonella henselae, responsible for cat scratch disease, which sometimes causes systemic symptoms. We report the case of an adolescent who came to the emergency room with intense myalgia, malaise, weight loss and splenomegaly. Blood tests showed high inflammatory markers. She had been in touch with a cat. Studies were carried out including: lower limbs MRI suggestive of bilateral inflammatory myositis, abdominal MRI with three previously undetected splenic lesions and dilated fundus examination that showed possible retinal arterial occlusion or vasculitis. After 21 days of intravenous antibiotic therapy (cefotaxime + cloxaciline), she became asymptomatic, but inflammatory markers remained high. Suspecting Bartonella henselaeinfection (myositis + chorioretinitis + splenic abscess), oral azithromycin and rifampicin were prescribed for 14 days. Blood tests and control MRI became normal, and IgG was positive.

Clonamiento, expresión y seroreactividad de la proteína recombinante de ensamblaje de lipopolisacáridos - D (LptD) de Bartonella bacilliformis / Cloning, expression and seroreactivity of the recombinant lipopolysaccharide assembly protein - D (LptD) from Bartonella bacilliformis

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.

Prevalence of Bartonella species in shelter cats and their ectoparasites in southeastern Brazil / Prevalência de espécies de Bartonella em gatos de abrigos e seus ectoparasitas no Sudeste do Brasil

Abstract Feline Bartonella can be transmitted to humans through cat scratches or bites, and between cats, by the flea Ctenocephalides felis. The study was carried out in order to investigate the occurrence of Bartonella DNA in cats living in shelters and their ectoparasites and the relationship between the infection status of cats and ectoparasites they host. Bartonella DNA was detected in 47.8% of the cat blood samples, 18.3% of C. felis fleas, 13.3% of flea egg pools and 12.5% of lice pools. B. henselae and B. clarridgeiae DNA were detected in cat fleas, while B. henselae, B. clarridgeiae and B. koehlerae were found in blood samples from bacteremic cats. Cats infested by positive ectoparasites showed approximately twice the odds of being infected. Our results indicate that shelter cats have high prevalence of Bartonella species that are known to be human pathogens. This highlights the importance of controlling infestations by ectoparasites to avoid cat and human infection.

Resumo Algumas espécies de Bartonella têm os felinos como principais hospedeiros reservatórios. Tais patógenos são transmitidos ao homem por intermédio da arranhadura ou mordedura de gatos e entre os gatos, por meio da pulga Ctenocephalides felis. O objetivo deste estudo foi investigar a ocorrência de DNA de Bartonella spp. em gatos de abrigos e seus ectoparasitas e a relação entre o estado de infecção dos gatos e dos ectoparasitas albergados por estes. Material genético bacteriano foi detectado em 47,8% das amostras de sangue de gatos, 18,3% das pulgas C. felis, 13,3% dos "pools" de ovos de pulgas e 12,5% dos "pools" de piolhos. DNA de B. henselae e B. clarridgeiae foi detectado em pulgas, e B. henselae, B. clarridgeiae e B. koehlerae, em amostras de sangue de gatos. Gatos infestados por ectoparasitas que carreavam DNA de Bartonella spp. demonstraram aproximadamente o dobro de chance de estarem infectados. Esses resultados indicam que os gatos de abrigos têm alta prevalência de infecção por espécies de Bartonella, capazes de causar doenças no homem. E também destacam a importância do controle e prevenção da infestação por ectoparasitas, no intuito de prevenir a infecção em gatos e humanos.

Chronic type 2 reaction possibly triggered by an asymptomatic Bartonella henselae infection in a leprosy patient

ABSTRACT As leprosy and leprosy reactions are the most prevalent infectious cause of physical disability, it is important to commit efforts to better understand these chronic reactions. Infections, even when asymptomatic, can trigger leprosy reactions and Bartonella spp. in turn, can cause chronic infections. We presented a case of a 51-year-old man who was admitted presenting with chronic type 2 leprosy reactions. He had a lepromatous form of leprosy that was histologically diagnosed six months after the onset of signs and symptoms compatible with a chronic type 2 reaction. He reported a history of a previous hepatitis B diagnosis. During a 24-month multidrug therapy (MDT), chronic reactions were partially controlled with prednisone and thalidomide. Thirty-three months following the leprosy treatment, he still experienced chronic reactions, and whole bacilli as well as globi were found on a new skin biopsy. Since coinfections can trigger type 2 reactions and the patient had close contact with animals and ticks, we investigated the presence of a Bartonella sp. infection. Bartonella henselae DNA was detected in a skin fragment obtained before the beginning of the leprosy retreatment. However, even after six months of a second leprosy MDT, he continued to experience type 2 chronic reactions. He was admitted to the hospital to undergo an intravenous antibiotic therapy for 14 days and then complete the treatment per os for ten more weeks. Leprosy reactions improved following the treatment for B. henselae. After completing the MDT treatment, he has been accompanied for sixty months with no signs of leprosy or leprosy reactions. The asymptomatic infection by B. henselaein this patient was considered the putative trigger of chronic leprosy reactions and leprosy relapse.

Two-year history of lymphadenopathy and fever caused by Bartonella henselae in a child

ABSTRACT We report the case of a 6-year-old boy that presented with enlarged lymph nodes on his neck. He complained of tiredness and discouragement, which worsened during feverish periods. There were no relevant laboratory test abnormalities and serological tests were not reactive. Bartonella henselae DNA was detected by species-specific nested polymerase chain reaction. After treatment, the patient progressed with no fever or lymphadenopathy. Bartonellosis is a group of infectious diseases caused by bacteria of the genus Bartonella. This case report is a useful reminder to clinicians that long-term fever of unknown origin can be related to B. henselae infection, even if the specific serology is not reactive.

Lymphedema secondary to bartonellosis as a differential diagnosis of breast cancer

In the presence of unilateral axillary lymphadenopathy associated with a breast radiological finding, breast cancer should constitute the main differential diagnosis. This fact is intensified when there is associated lymphedema. We present a case of a patient in these conditions, for whom breast cancer was not confirmed, and a subsequent evaluation showed that it was cat-scratch disease. This report constitutes the second case of association between lymphedema and bartollenosis.

Serological review of Bartonella henselae and Bartonella quintana infection among Malaysian patients with unknown causes of febrile illnesses

@#Limited information is available on human exposure to Bartonella infection, i.e., Bartonella henselae (causative agent of cat scratch disease) and Bartonella quintana (causative agent of trench fever) in West Malaysia. This study reports a review of serological findings obtained from patients attending to a teaching hospital in Klang Valley, Malaysia. An indirect immunofluorescence assay (IFA) was used to determine IgG and IgM antibody titers against B. henselae and B. quintana. In a pilot study conducted between 2013-2015, IgG antibodies against Bartonella spp. (either B. quintana and B. henselae) were detected in 14 (36.8%) of 38 patients who were clinically suspected of rickettsial infections, while IgM antibody was detected in 4 (10.5%) patients. This has prompted us to investigate the serologic responses of patients who were clinically suspected of other febrile causes besides rickettsial infection. Of the 59 serum samples analysed in a follow-up investigation, Bartonella IgG antibodies were detected from 7 (11.9%) patients, of which 5 (27.8%) and 2 (18.2%) patients were clinically suspected of rickettsial infection (n=18) and dengue (n=11), respectively. None of the sera obtained from the leptospirosis (n=10), legionellosis (n=10) and mycoplasma infection (n=10) groups were seropositive to Bartonella spp. The review of Bartonella serological findings in this study highlights that Bartonella infection is not uncommon and should be considered as one of the causes for febrile illness in Malaysia.

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

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

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