Presence of intracellular bacterial communities in uroepithelial cells, a potential reservoir in symptomatic and non-symptomatic people.

BACKGROUND: Urinary tract infection is one of the most common infections in humans, affecting women in more proportion. The bladder was considered sterile, but it has a urinary microbiome. Moreover, intracellular bacteria (IB) were observed in uroepithelial cells from children and women with urinary tract infections (UTIs). Here, we evaluated the presence of IB in urine from healthy people and patients with UTI symptoms. METHODS: Midstream urine was self-collected from 141 donors, 77 females and 64 males; 72 belonged to the asymptomatic group and 69 were symptomatic. IB was characterized by a culture-dependent technique and visualized by confocal microscopy. Urine was also subjected to the classical uroculture and isolated bacteria were identified by MALDI-TOF. RESULTS: One-hundred and fifteen uroculture were positive. A significant association was observed between the presence of symptoms and IB (P = 0.007). Moreover, a significant association between the presence of IB, symptoms and being female was observed (P = 0.03). From the cases with IB, Escherichia coli was the most frequent microorganism identified (34.7%), followed by Stenotrophomonas maltophilia (14.2%), Staphylococcus spp (14.2%), and Enterococcus faecalis (10.7%). Intracellular E. coli was associated with the symptomatic group (P = 0.02). Most of the intracellular Staphylococcus spp. were recovered from the asymptomatic group (P = 0.006). CONCLUSIONS: Intracellular bacteria are present in patients with UTI but also in asymptomatic people. Here, we report for the first time, the presence of S. maltophilia, Staphylococcus spp., and Enterobacter cloacae as intracellular bacteria in uroepithelial cells. These findings open new insights into the comprehension of urinary tract infections, urinary microbiome and future therapies. Uroculture as the gold standard could not be enough for an accurate diagnosis in recurrent or complicated cases.

In house-development of a rapid immunochromatographic test for the detection of Escherichia coli in urine samples.

Urinary tract infections (UTIs) are a common health concern. Urine culture is the "gold standard" for UTI diagnosis but takes 48h. Rapid methods like dipstick tests are used as point-of-care tests. However, their sensitivity and specificity are variable. In this work, a rapid immunochromatographic test (IT) for detecting Escherichia coli in urine was developed, and its performance was evaluated in urine samples from patients with suspected UTI. The "universal lateral flow assay kit" was employed using an E. coli capture antibody. One hundred and five (105) urine samples were analyzed using the IT, dipstick test, and urine culture. The sensitivity of the IT was 74.5%, specificity 88.9%, positive predictive value (PPV) 86.3%, and negative predictive value (NPV) 78.7%. The combination of the IT with the dipstick test increases sensitivity to 94.1%, specificity to 66.7%, PPV to 72.7%, and NPV to 92.3%. Using the IT for detecting E. coli in urine could be a valuable technique for UTI screening, showing better specificity and diagnostic precision but lower sensitivity than the dipstick test. Based on these results, we propose that the combined use of both screening techniques would allow a rapid and more precise diagnosis of UTI, rationalizing the indication for empirical antibiotics.

Gold-, silver- and magnesium-doped zinc oxide nanoparticles prevents the formation of and eradicates bacterial biofilms.

Aim: This work aimed to synthesize magnesium-doped zinc oxide, silver and gold nanoparticles (Nps) and to evaluate their potential to prevent and eradicate Escherichia coli, Proteus mirabilis, Staphylococcus aureus, Acinetobacter baumannii and Pseudomonas aeruginosa biofilms. Materials & methods: The Nps were synthesized by precipitation and metallic reduction techniques. Physicochemical and biological characterization of Nps was performed. Results: All the Nps tested were able to inhibit the formation of E. coli, P. mirabilis, S. aureus and A. baumannii biofilms. The effects on the eradication of preformed biofilms were variable, although all the Nps tested were able to eradicate A. baumannii biofilms. Conclusion: The observed effects make the Nps suitable for coating surfaces and/or antibiotic carriers with medical interest.

Antibiotic susceptibility and fosfomycin resistance characterization in a cohort of children older than 6 years of age with urinary tract infection.

Fosfomycin tromethamol (FT) was reintroduced as an option for the treatment of low urinary tract infection (UTI) in children. In this study, we described the antibiotic sensitivity and mechanisms of resistance to fosfomycin in isolates from children older than 6 years with UTI. Urine culture and antibiotic susceptibility study were performed. In fosfomycin resistant strains, PCR for fos, blaCTX-M was performed followed by classification by phylogenetic group and sequencetyping. Escherichia coli was the most frequent etiological agent (89.2%). The susceptibility percentages were: fosfomycin 97.9%; amoxicillin-clavulanate 92.7%; cefuroxime and ceftriaxone 99%; nitrofurantoin 94.4%. An E. coli strain (ST69, phylogenetic group D) was resistant to fosfomycin (MIC 256mg/l) and carried the blaCTX-M-14 and fosA3 genes in a 45kb IncN-type plasmid. This is the first report of E. coli ST69 with blaCTX-M-14/fosA3 of human origin.

Fosfomicina en el tratamiento de la infección urinaria baja en niños mayores de 6 años. Evolución clínico-microbiológica / Fosfomycin for low urinary-tract infections in children of over 6 years of age. Clínical-microbiological evolution / Fosfomicina no tratamento da infecção do trato urinário baixo em crianças maiores de 6 anos de idade. Evolução clínico-microbiológica

Resumen: Introducción: fosfomicina trometamol (FT) representa una alternativa al tratamiento de la infección del tracto urinario (ITU) baja. Uruguay no dispone de información acerca de su uso en niños. Objetivo: describir la evolución clínica y microbiológica de una cohorte de niños mayores de 6 años con ITU baja tratados con FT. Material y método: se incluyeron niños mayores de 6 años con ITU baja de dos prestadores de salud de Montevideo, entre 1/2/2018 - 30/6/2019. A todos se indicó FT 2 g monodosis y urocultivo de control. Se realizó seguimiento telefónico. Se evaluó: clínica, antecedentes de ITU, microorganismo, susceptibilidad antimicrobiana y evolución: tiempo de resolución clínica, resolución microbiológica, efectos adversos, recurrencia en los primeros tres meses. Resultados: se incluyeron 46 niños, mediana de edad 9,4 años, antecedentes de ITU 13. Presentaron disuria 44, tenesmo 33, polaquiuria 31. Microorganismo aislado: E. coli 43, S. saprophyticus 2, Proteus sp 1. Todos susceptibles a FT, excepto S. saprophyticus naturalmente resistente. Resolución clínica en 48 horas: 42. Se obtuvo urocultivo de control en 31/46 niños: resolución microbiológica 22, no resolución 5 y contaminado 4. Presentaron efectos adversos 9: vómitos 1, diarrea 8 y cefalea 1. Seguimiento telefónico a 40/46 pacientes: reinfecciones al mes de tratamiento: 6. Conclusiones: no se registró resistencia adquirida en los microorganismos. Se observó resolución clínica en las primeras 48 horas en la mayoría de los casos. Los efectos adversos fueron leves. Ocurrieron reinfecciones en una proporción pequeña. Los resultados avalan a FT como alternativa terapéutica para ITU baja en mayores de 6 años.

Summary: Introduction: fosfomycin tromethamine (FT) is an alternative to the treatment of low urinary tract infection (UTI). Uruguay does not have information about its use in children. Objective: to describe the clinical and microbiological evolution of a cohort of children older than 6 years of age with low UTI treated with FT. Materials and methods: we included children of over 6 years of age with low UTI from two health providers in Montevideo between 2/1/2018 and 6/30/2019. We prescribed a single dose of FT 2 g and a control urine culture to all patients. We carried out a telephone follow-up and assessed their clinical record, history of UTI, microorganisms, antimicrobial susceptibility and evolution: time of clinical resolution, microbiological resolution, adverse effects, and recurrence during the first 3 months. Results: 46 children were included, median age 9.4 years, history of UTI 13. 44 presented dysuria, 33 tenesmus, 31 pollakiuria. Isolated microorganism: E. coli 43, S. saprophyticus 2, Proteus sp 1. All susceptible to FT, except S. saprophyticus, naturally resistant. Clinical resolution in 48 hours: 42. Control urine culture was obtained in 31/46 children: microbiological resolution 22, no resolution 5 and contaminated 4. Adverse effects 9: vomiting 1, diarrhea 8, and headache 1. Telephone follow-up carried out for 40 / 46 patients: reinfections after one month of treatment: 6. Conclusions: microorganisms had not acquired resistance. Most cases showed clinical resolution during the first 48 hours. Adverse effects were mild. Reinfections occurred in a small proportion. The results support FT as a therapeutic alternative for low UTI for the case of children of over 6 years of age.

Resumo: Introdução: A fosfomicina trometamina (FT) é uma alternativa ao tratamento da infecção do trato urinário baixo (ITU). O Uruguai não possui informações sobre seu uso em crianças. Objetivo: Descrever a evolução clínica e microbiológica de uma coorte de crianças maiores de 6 anos de idade com ITU baixa tratada com TF. Materiais e métodos: Foram incluídas crianças maiores de 6 anos de com ITU baixa de dois provedores de saúde em Montevidéu; no período 1/2 / 2018 e 30/06/2019. Todos os pacientes receberam indicação de FT 2 g em dose única, cultura de urina e controle. Realizou-se um rastreamento por telefone. Se avaliou: prontuário clínico, história de ITU, microrganismos, suscetibilidade a antimicrobianos e evolução: tempo de resolução clínica, resolução microbiológica, efeitos adversos, recorrência nos primeiros 3 meses. Resultados: Incluíram-se 46 crianças, mediana de idade 9,4 anos, história de ITU 13. 44 delas apresentaram disúria, tenesmo 33, polaciúria 31. Microrgoanismo isolado: E. coli 43, S. saprophyticus 2, Proteus sp 1. Todas suscetíveis a FT, exceto S. saprophyticus, naturalmente resistente. Resolução clínica em 48 horas: 42. Obtivemos cultura de urina controle em 31/46 crianças: resolução microbiológica 22, sem resolução 5 e contaminada 4. 9 delas apresentaram efeitos adversos 9: vômito 1, diarreia 8 e dor de cabeça 1. Realizamos acompanhamento telefônico em 40 / 46 pacientes: reinfecções um mês após tratamento, 6. Conclusões: Os microrganismos não adquiriram resistência. Na maioria dos casos observou-se resolução clínica nas primeiras 48 horas. Os efeitos adversos foram leves. As reinfecções ocorreram em pequena proporção. Os resultados apoiam o TF como uma alternativa terapêutica para ITU baixa para casos de crianças maiores de 6 anos de idade.

Selection of Effective Antibiotics for Uropathogenic Escherichia coli Intracellular Bacteria Reduction.

Urinary tract infections (UTI) are one of the most frequent bacterial infections in humans, being Uropathogenic Escherichia coli (UPEC), the most common etiological agent. The ability of UPEC to invade urothelial cells and to form intracellular bacterial communities (IBC) has been described. Therefore, UPEC can persist in the urinary tract producing recurrent infections, resisting antibiotic activity. The objective of the present work was to analyze the ability of a collection of UPEC clinical isolates to invade bladder epithelial cells in vitro and the activity of different classes of antibiotics on intracellular bacteria. We selected 23 UPEC clinical isolates that had been previously detected intracellularly in desquamated bladder epithelial cells from patients' urine. A cellular invasion assay using the T24 bladder cell line was used. Intracellular bacteria was confirmed by laser confocal microscopy. All the strains were able to invade the cells with different percentages of intracellular bacterial survival (0.7 to 18%). However, no significant relationship was found between the percentage of in vitro infection and the presence of IBC in desquamated urine cells. In vitro, intracellular bacteria were confirmed in four representative strains by confocal laser microscopy. Ceftriaxone, ciprofloxacin and, azithromycin in vitro activity on intracellular bacteria were evaluated. Amikacin was used as a negative control. All the antibiotics tested, except amikacin, significantly decreased the number of intracellular bacteria. Ciprofloxacin was the antibiotic that induced the highest decrease percentage. Conclusions: All UPEC clinical isolates could invade bladder epithelial cells in vitro. Ceftriaxone, ciprofloxacin, and azithromycin can reduce the percentage of intracellular bacteria in vitro. In vivo studies are needed to confirm the utility of these antibiotics for intracellular bacteria reduction in UTI.

Interpretación de los tests de sospecha e inicio de la terapia antibiótica empírica en infecciones urinarias / How to interpret suspicion tests and when to start empirical antibiotic therapy for urinary tract infections / Interpretação dos testes de suspeita e início da antibioticoterapia empírica no caso das infecções urinárias

Resumen: Introducción: las infecciones del tracto urinario (ITU) son motivo de consulta frecuente. Los nitritos y la estearasa leucocitaria (EL) o leucocitos en orina son marcadores de sospecha, pero su presencia es variable. Objetivo: determinar la validez de los tests de sospecha y describir la indicación de antibióticos en función de los resultados. Material y método: estudio descriptivo, retrospectivo, de niños con sospecha de ITU estudiados con nitritos, EL y urocultivo entre 2016 y 2018. Resultados: se estudiaron 137 pacientes por sospecha de ITU, confirmándose mediante urocultivo en 27 (19,7%). En niños de 1 mes a 3 años, el principal motivo de sospecha de ITU fue fiebre sin foco y la sensibilidad de leucocitos/EL fue de 75%, especificidad 65,6%, valor predictivo positivo (VPP) 35,2% y valor predictivo negativo (VPN) 91,3%. En el caso de los nitritos y EL la sensibilidad fue 43,7%, especificidad 93,7%, VPP 63,6% y VPN 87%. En mayores de 3 años la fiebre, junto a síntomas urinarios, fue el motivo de sospecha de ITU más frecuente, pero con baja sensibilidad diagnóstica. La sensibilidad de los leucocitos fue 72,7%, especificidad 72,9%, VPP 38,1% y VPN 92,1%. En leucocitos y nitritos la sensibilidad fue de 63,6%, especificidad 93,7%, VPP 70% y VPN 91,8%. El 65% de los niños mayores de 1 mes con sospecha de ITU recibieron antibióticos empíricamente. Se confirmó ITU solo en 29,6% de los que recibieron antibiótico. Conclusiones: existe sobrediagnóstico de ITU en pediatría. La baja sensibilidad y especificidad de los síntomas y los tests de sospecha conllevan a un uso irracional de antimicrobianos.

Summary: Introduction: urinary tract infections (UTIs) are a cause of frequent consultation. Nitrites and leukocyte esterase (LE) or leukocytes in urine are suspected markers, but their presence is variable. Objective: determine the validity of the suspicion tests and describe the antibiotic prescription based on the results. Materials and methods: descriptive, retrospective study of children with suspected UTIs analyzed with nitrites, EL and urine culture between 2016 and 2018. Results: 137 patients were studied for suspected UTIs, and they were confirmed through urine cultures in 27 (19.7%). The main reason for suspected UTIs was fever of unknown origin (FUO) for children aged one month to 3 years of age, and leukocyte / LE sensitivity was 75%, specificity 65.6%, PPV 35.2% and NPV 91.3%. In the case of nitrites and LE the sensitivity was 43.7%, specificity 93.7%, PPV 63.6% and NPV 87%. For children of over 3 years of age, fever and urinary symptoms were the most frequent suspicion of UTI, but they showed low diagnostic sensitivity. Leukocyte sensitivity was 72.7%, specificity 72.9%, PPV 38.1% and NPV 92.1%. For leukocytes and nitrites, the sensitivity was 63.6%, specificity 93.7%, PPV 70% and NPV 91.8%. 65% of children of over 1 month of age with suspected UTI received empirical antibiotics. ITUs were confirmed in only 29.6% of those who received antibiotics. Conclusions: pediatric UTIs are over diagnosed. The symptoms' low sensitivity and specificity as wekk as suspicion tests lead to an excessive use of antimicrobials.

Resumo: Introdução: as infecções do trato urinário (ITU) são causa de consultas frequentes. Nitritos e esterase de leucócitos (EL) ou leucócitos na urina são os marcadores suspeitos, mas sua presença é variável. Objetivo: determinar a validade dos testes de suspeita e descrever a prescrição de antibióticos com base nos resultados. Materiais e métodos: estudo descritivo, retrospectivo, de crianças com suspeita de ITU através de nitritos, EL e cultura de urina entre 2016-2018. Resultados: 137 pacientes foram analisados por suspeita de ITU e confirmou-se a cultura de urina em 27 deles (19,7%). Em crianças de 1 mês a 3 anos, o principal motivo para suspeita de ITU foi febre sem foco e sensibilidade a leucócitos / EL foi de 75%, especificidade de 65,6%, PPV 35,2% e VPN 91,3%. No caso de nitritos e EL, a sensibilidade foi de 43,7%, especificidade de 93,7%, VPP de 63,6% e VPN de 87%. No caso de crianças de mais de três anos de idade, a febre e sintomas urinários foram as suspeitas mais frequentes de ITU, mas com baixa sensibilidade diagnóstica. A sensibilidade dos leucócitos foi de 72,7%, especificidade de 72,9%, PPV 38,1% e VPN de 92,1%. A sensibilidade dos leucócitos e nitritos foi de 63,6%, especificidade de 93,7%, PPV 70% e VPN 91,8%. 65% das crianças acima de 1 mês de idade com suspeita de ITU receberam antibióticos empiricamente. A UIT foi confirmada em apenas 29,6% daqueles que receberam antibióticos. Conclusões: há um excessivo diagnóstico de ITU em pediatria. A baixa sensibilidade e especificidade dos sintomas e testes de suspeita levam a um uso irracional de antimicrobianos.

Magnesium-doped zinc oxide nanoparticles alter biofilm formation of Proteus mirabilis.

Aim:Proteus mirabilis biofilms colonize medical devices, and their role in microbial pathogenesis is well established. Magnesium-doped zinc oxide nanoparticles (ZnO:MgO NPs) have potential antimicrobial properties; thus, we aimed at evaluating the antibiofilm activity of ZnO:MgO NPs against P. mirabilis biofilm. Materials & methods: After synthesis and characterization of ZnO:MgO NPs and their addition to a polymer film, we evaluated the stages of P. mirabilis biofilm development over glass coverslip covered by different concentrations of ZnO:MgO NPs. Results: Low concentrations of ZnO:MgO NPs affect the development of P. mirabilis biofilm. Descriptors showed reduced values in bacterial number, bacterial volume and extracellular material. Conclusion: Our results highlight this new application of ZnO:MgO NPs as a potential antibiofilm strategy in medical devices.

Fosfomycin tromethamine activity on biofilm and intracellular bacterial communities produced by uropathogenic Escherichia coli isolated from patients with urinary tract infection.

Fosfomycin tromethamine (FT), an old antibiotic revived as a new strategy to overcome antibiotic resistance, is an excellent option for the treatment of lower urinary tract infection (UTI). During UTI, Escherichia coli produces biofilms and could invade the bladder epithelial cells, developing intracellular bacterial communities (IBC). The present work aimed to evaluate the activity of FT on biofilms and IBC from clinical isolates of E. coli. A total of 38 E. coli clinical UTI isolates previously characterized as biofilm and IBC producers were studied. FT susceptibility was evaluated and its activity on 48 h biofilm was determined by microtiter plate-based biofilm assay comparing three different antibiotic concentrations. Two UPEC strains were selected to evaluate FT activity on IBC in vitro using T24 bladder cells. The survival percentage of intracellular bacteria after 24 h exposure to FT was calculated and compared to the percentage of intracellular bacteria without antibiotic. All the strains were susceptible to FT. FT produced a significant reduction of biofilms at the three concentrations tested, compared to the control. However, no statistically effect on IBC was observed after 24 h of fosfomycin exposure in cell culture. FT is a good option for bacterial biofilm reduction within UTI. However, it does not affect IBC.

Effect of different antibiotics on biofilm produced by uropathogenic Escherichia coli isolated from children with urinary tract infection.

Recurrent urinary tract infections (UTIs) occur frequently in children and women. Intracellular bacterial communities (IBCs) and biofilm formation by Escherichia coli are risk factors for recurrence. The aim of this study was to evaluate the effect of different antibiotics on biofilms by E. coli strains isolated from children with UTI and to correlate virulence factors and IBCs with biofilm formation. A total of 116 E. coli strains were tested for biofilm formation using the crystal violet microplate technique. 58.6% of the strains did not produce biofilm, while 16.4%, 18.1% and 6.8% formed weak, moderate and strong biofilms, respectively. No correlation was found between the ability to form biofilms and the presence of IBCs. Biofilm formation was significantly associated with pili P codifying genes, whereas other virulence factors were not statistically associated. Antibiotics, including ampicillin, cephalothin, ceftriaxone, ceftazidime, amikacin and ciprofloxacin, were evaluated at different concentrations after 48 h of biofilm formation. Except ampicillin, the other antibiotics tested induced a significant reduction of biofilm biomass. In the case of recurrent UTIs potentially associated with the presence of biofilm, the use of third-generation cephalosporin, fluoroquinolones and aminoglycosides could be recommended. These antibiotics demonstrated to reduce biofilm biomass produced even by resistant strains.

Enteropathogens associated with acute diarrhea in children from households with high socioeconomic level in uruguay.

Infectious diarrhea, a common disease of children, deserves permanent monitoring in all social groups. To know the etiology and clinical manifestations of acute diarrhea in children up to 5 years of age from high socioeconomic level households, we conducted a descriptive, microbiological, and clinical study. Stools from 59 children with acute community-acquired diarrhea were examined, and their parents were interviewed concerning symptoms and signs. Rotavirus, adenovirus, and norovirus were detected by commercially available qualitative immunochromatographic lateral flow rapid tests. Salmonella, Campylobacter, Yersinia, and Shigella were investigated by standard bacteriological methods and diarrheagenic E. coli by PCR assays. We identified a potential enteric pathogen in 30 children. The most frequent causes of diarrhea were enteropathogenic E. coli (EPEC), viruses, Campylobacter, Salmonella, and Shiga-toxin-producing E. coli (STEC). Only 2 patients showed mixed infections. Our data suggest that children with viral or Campylobacter diarrhea were taken to the hospital earlier than those infected with EPEC. One child infected with STEC O26 developed "complete" HUS. The microbiological results highlight the importance of zoonotic bacteria such as atypical EPEC, Campylobacter, STEC, and Salmonella as pathogens associated with acute diarrhea in these children. The findings also reinforce our previous communications about the regional importance of non-O157 STEC strains in severe infant food-borne diseases.

Intracellular bacteria in the pathogenesis of Escherichia coli urinary tract infection in children.

BACKGROUND: Uropathogenic Escherichia coli (UPEC) is the most common agent of urinary tract infection (UTI). The classic model of pathogenesis proposes the ascent of UPEC by the urethra and external adherence to the urothelium. Recently, the ability of UPEC to invade urothelial cells and to form intracellular bacterial communities (IBCs) has been described. METHODS: The objective of the present study was to determine the presence of intracellular bacteria (IB) in children with UTI caused by E. coli and to characterize its virulence attributes and its relation with clinical outcomes. One hundred thirty-three children with E. coli UTI who attended a reference children's hospital between June and November 2012 were included. Urine samples were analyzed by optical and confocal microscopy looking for exfoliated urothelial cells with IB. Phylogenetic group and 24 virulence factors of UPEC were determined using multiplex polymerase chain reaction. Medical records were analyzed. RESULTS: The presence of IB was detected in 49 of 133 (36.8%) samples by confocal microscopy, in 30 cases as IBC, and in 19 as isolated intracellular bacteria (IIB). Only 50% of these cases could be detected by light microscopy. Seventy-four medical records were analyzed, 34 with IBC/IIB, 40 without IB. Any virulence gene was associated with IBC/IIB. The presence of IBC/IIB was associated with recurrent UTI (odds ratio [OR], 3.3; 95% confidence interval [CI], 1.3-9; P = .017), especially in children without urinary tract functional or morphological abnormalities (OR, 8.0; 95% CI, 2.3-27.4; P = .000). IBCs were associated with lower urinary tract syndrome (OR, 3.6; 95% CI, 1.1-11.8; P = .05) and absence of fever (P = .009). CONCLUSIONS: IBCs/IIB could explain a high proportion of children with recurrent UTI.

Urinary tract infection in Uruguayan children: Aetiology, antimicrobial resistance and uropathogenic Escherichia coli virulotyping.

Uropathogenic Escherichia coli (UPEC) is the most frequent cause of urinary tract infection (UTI). Virulence factors (VFs) of UPEC in children are not well known. Circulating antibiotic resistance mechanisms in the community are increasing. In this study, the aetiological agents of UTI and antibiotic resistance mechanisms of 124 strains isolated from urine cultures from children with community-acquired UTI were determined. Virulotyping of isolated E. coli strains was also described. ß-Lactam, fluoroquinolone and sulfonamide resistance genes as well as integrons were detected by PCR. E. coli phylogenetic groups and 25 VFs were sought by multiplex PCR. E. coli was the most frequent aetiological agent (88.7%), of which 48.2% belonged to phylogenetic group D and 35.5% to group B2. Moreover, 81.8% were considered UPEC and >93% had virulence structures, with kpsMTII, fimH and iutA being the most frequent. Most of the E. coli isolates were susceptible to amoxicillin/clavulanic acid (AMC) (87.3%), nitrofurantoin (97.3%), cefuroxime and third-generation cephalosporins (100%). Resistance levels to oxyimino-cephalosporins were higher in non-E. coli isolates, with circulation of integrons, blaCTX-M-2 and blaCMY-2 detected in the community. Moreover, 8.1% of isolates were resistant to fluoroquinolones, with qnrB found in two isolates. Resistance to trimethoprim/sulfamethoxazole was found in 37.9% of isolates, with 85.5% harbouring sul genes. E. coli isolated from children with UTI presented high rates of VFs. Nitrofurantoin, AMC and cefuroxime would be suitable antibiotics to treat UTI in children. However, the presence of integrons (fundamentally class 1) and circulation of broad-spectrum ß-lactamases in the community makes continuous surveillance necessary.

[Neonatal cholestasis due to congenital toxoplasmosis. Case report]. / Colestasis neonatal por toxoplasmosis congénita. A propósito de un caso clínico.

Neonatal cholestasis is the manifestation of many different diseases. Its early etiological diagnosis is crucial, since treatment before 60 days of life changes the prognosis in children with biliary atresia. Congenital toxoplasmosis can be asymptomatic in the newborn, or have mainly neurological, ophthalmological or gastrointestinal symptoms (hepatomegaly, cholestatic jaundice). Neonatal cholestasis secondary to congenital toxoplasmosis is not a situation frequently reported. We report the case of an infant with neonatal cholestasis due to a congenital toxoplasmosis, in order to discuss the difficulties in establishing the etiological diagnostic and to review the indications of invasive studies such as liver biopsy in these situations.

Colestasis neonatal por toxoplasmosis congénita: A propósito de un caso clínico / Neonatal cholestasis due to congenital toxoplasmosis: Case report

La colestasis neonatal es la forma de presentación de diversas enfermedades; es necesario un diagnóstico etiológico temprano, ya que el tratamiento antes de los 60 días de vida cambia el pronóstico en los niños que presentan atresia biliar. La toxoplasmosis congénita puede ser asintomática en el recién nacido, o presentar fundamentalmente alteraciones neurológicas, oftalmológicas y hepáticas (hepatomegalia, ictericia no colestásica). La colestasis neonatal secundaria a toxoplasmosis congénita no es una situación informada con frecuencia. Se presenta el caso de un lactante con colestasis neonatal cuya etiología responde a una toxoplasmosis congénita, con el objetivo de discutir las difcultades en establecer el diagnóstico etiológico y las indicaciones de realizar estudios invasivos, como la biopsia hepática, en estas situaciones.(AU)

Neonatal cholestasis is the manifestation of many different diseases. Its early etiological diagnosis is crucial, since treatment before 60 days of life changes the prognosis in children with biliary atresia. Congenital toxoplasmosis can be asymptomatic in the newborn, or have mainly neurological, ophthalmological or gastrointestinal symptoms (hepatomegaly, cholestatic jaundice). Neonatal cholestasis secondary to congenital toxoplasmosis is not a situation frequently reported. We report the case of an infant with neonatal cholestasis due to a congenital toxoplasmosis, in order to discuss the diffculties in establishing the etiological diagnostic and to review the indications of invasive studies such as liver biopsy in these situations.(AU)

Colestasis neonatal por toxoplasmosis congénita: A propósito de un caso clínico / Neonatal cholestasis due to congenital toxoplasmosis: Case report

La colestasis neonatal es la forma de presentación de diversas enfermedades; es necesario un diagnóstico etiológico temprano, ya que el tratamiento antes de los 60 días de vida cambia el pronóstico en los niños que presentan atresia biliar. La toxoplasmosis congénita puede ser asintomática en el recién nacido, o presentar fundamentalmente alteraciones neurológicas, oftalmológicas y hepáticas (hepatomegalia, ictericia no colestásica). La colestasis neonatal secundaria a toxoplasmosis congénita no es una situación informada con frecuencia. Se presenta el caso de un lactante con colestasis neonatal cuya etiología responde a una toxoplasmosis congénita, con el objetivo de discutir las difcultades en establecer el diagnóstico etiológico y las indicaciones de realizar estudios invasivos, como la biopsia hepática, en estas situaciones.

Neonatal cholestasis is the manifestation of many different diseases. Its early etiological diagnosis is crucial, since treatment before 60 days of life changes the prognosis in children with biliary atresia. Congenital toxoplasmosis can be asymptomatic in the newborn, or have mainly neurological, ophthalmological or gastrointestinal symptoms (hepatomegaly, cholestatic jaundice). Neonatal cholestasis secondary to congenital toxoplasmosis is not a situation frequently reported. We report the case of an infant with neonatal cholestasis due to a congenital toxoplasmosis, in order to discuss the diffculties in establishing the etiological diagnostic and to review the indications of invasive studies such as liver biopsy in these situations.

[Treatment failure due to acquisition of antibiotic resistance: report of two clinical cases]. / Fallo terapéutico por resistencia antibacteriana intra-tratamiento, a propósito de dos casos clínicos.

OBJECTIVE: We describe two cases of treatment failure due to intra-treatment acquisition of antibiotic resistant microorganisms with the aim of highlighting the possible molecular mechanisms by which treatment failure occurred. PATIENTS AND METHODS: We analyzed the clinical histories and the isolates obtained from 2 patients, one with a urinary tract infection (UTI) by E. coli, initially treated with cefuroxim (to which the isolate was susceptible), and another with osteoarthritis (OA) treated initially with meropenem plus vancomycin, developing K. pneumoniae susceptible to meropenem. During treatment, in both patients, resistant microorganisms were isolated, and empirical therapy was modified, initially with ceftriaxone and afterwards meropenem in case 1, and adding amikacin in case 2. Both strains (per patient) were compared by PFGE and resistance genes were sought by PCR. RESULTS: Regarding the UTI, the initial strain acquired an IncFIB SHV-5-producing plasmid. In the OA case, the initial susceptible strain was substituted by a CTX-M-9 and AadB-AadA2-Aac(6')Ib-producing K. pneumoniae.

Detection of intracellular bacterial communities in a child with Escherichia coli recurrent urinary tract infections.

The formation of intracellular bacterial communities (IBC) has been proposed as a new pathogenic model for urinary tract infections. Scarce reports describe this phenomenon in humans. We describe the presence of IBC in uroepithelial cells of a child with recurrent urinary infections. Urine specimen was collected from a child with Escherichia coli UTI and analyzed by light and confocal laser scanning microscopy (CLSM). The capability of this strain to produce intracellular infection in bladder tissue was confirmed in mice models. Escherichia coli phylogenetic group, presence of virulence factors genes, and its multiple locus sequence type were determined. CLSM showed large collections of morphologically coccoid and rod bacteria in eukaryotic cells cytoplasm, even seemingly protruding from the cells. Escherichia coli EC7U, ST3626, harbored type 1, P, and S/F1C fimbriae and K1 capsule genes. In this report, we confirm the presence of IBC in children with UTI, as it has been described before in women.