Current Status of Antimicrobial Stewardship Programs in São Paulo Hospitals

OBJECTIVES: Antimicrobial stewardship programs (ASPs) comprise coordinated interventions designed to improve antimicrobial use. Understanding the current structure of ASP hospitals will support interventions for the improvement of these programs. This study aimed to describe the status of ASPs in hospitals in São Paulo, Brazil. METHODS: A cross-sectional survey was conducted on the ASPs of hospitals in the state of São Paulo from March to July 2018. Through interviews by telephone or e-mail, we queried which components of the Infectious Diseases Society of America/Society for Healthcare Epidemiology of America and Centers for Disease Control and Prevention guidelines were implemented. RESULTS: The response rate was 30% (28/93 hospitals), and 26 hospitals (85%) reported having a formal ASP. The most frequently implemented strategies were antimicrobial surgical prophylaxis guidelines (100%), empiric sepsis guidelines (93%), and the presence of ASP team members during bedside rounds (96%). The least commonly implemented strategies included prior authorization for all antimicrobials (11%), pharmacokinetic monitoring, and an adjustment program for patients on IV aminoglycosides (3%). Regarding the metrics of the ASP, the most common indicator was the rate of antimicrobial resistance (77%). Eighteen hospitals evaluated antimicrobial consumption using defined daily dose, and only 29% evaluated the days of therapy; 61% of hospitals reported their results to the hospital administration and 39% to the prescribers. CONCLUSIONS: Most hospitals have a formal and active ASP, but with timely actions. We observed inconsistencies between what program leaders understand as the main objective of ASP and the metrics used to evaluate it. Part of the effort for the next few years should be to improve program evaluation metrics and to provide feedback to physicians and hospital leadership

Polymerase chain reaction targeting 16S ribosomal RNA for the diagnosis of bacterial meningitis after neurosurgery

OBJECTIVES: Bacterial and aseptic meningitis after neurosurgery can present similar clinical signs and symptoms. The aims of this study were to develop and test a molecular method to diagnose bacterial meningitis (BM) after neurosurgery. METHODS: A 16S ribosomal RNA gene PCR-based strategy was developed using artificially inoculated cerebrospinal fluid (CSF) followed by sequencing. The method was tested using CSF samples from 43 patients who had undergone neurosurgery and were suspected to suffer from meningitis, and from 8 patients without neurosurgery or meningitis. Patients were classified into five groups, confirmed BM, probable BM, possible BM, unlikely BM, and no meningitis. RESULTS: Among the samples from the 51 patients, 21 samples (41%) were culture-negative and PCR-positive. Of these, 3 (14%) were probable BM, 4 (19%) were possible BM, 13 (62%) were unlikely BM, and 1 (5%) was meningitis negative. Enterobacterales, non-fermenters (Pseudomonas aeruginosa and Acinetobacter baumannii), Staphylococcus haemolyticus, Granulicatella, Variovorax, and Enterococcus cecorum could be identified. In the group of patients with meningitis, a good agreement (3 of 4) was observed with the results of cultures, including the identification of species. CONCLUSION: Molecular methods may complement the diagnosis, guide treatment, and identify non-cultivable microorganisms. We suggest the association of methods for suspected cases of BM after neurosurgery, especially for instances in which the culture is negative

Statewide evaluation of infection control measures for preventing coronavirus disease 2019 in hemodialysis facilities

OBJECTIVE: This study aimed to evaluate the occurrence of coronavirus disease 2019 (COVID-19) in hemodialysis facilities and the occurrence of and risk factors for clustering of COVID-19 cases. METHODS: We conducted a cross-sectional online survey between March and July 2020, in all dialysis facilities in São Paulo state, using Google Forms. The online questionnaire contained questions addressing specific components of infection prevention and control practices and the number of cases during the COVID-19 pandemic. RESULTS: A total of 1,093 (5%) COVID-19 cases were reported among 20,984 patients; approximately 56% of the facilities had ≥1 cluster. Most facilities implemented various measures (such as allocation of dedicated COVID-19 areas/shifts, symptom screening, environmental disinfection, and maintenance of adequate ventilation) to prevent the transmission of severe acute respiratory syndrome coronavirus 2. Clustering of COVID-19 cases was suspected in only 7% of dialysis facilities. The only variable associated with this event was the performance of aerosol-generating procedures (odds ratio: 4.74; 95% confidence interval: 1.75-12.86). CONCLUSION: Attention should be paid to avoiding the performance of aerosol-generating procedures in dialysis facilities and monitoring the clustering of cases

Streptococcus constellatus causing concomitant extra and intracranial abscesses complicated with sagittal sinus thrombosis

Streptococcus constellatus is a gram-positive coccus member of the Streptococcus anginosus group (SAG). It can be found in the oral flora, and may cause abscess more commonly in the gastrointestinal tract, lungs, and heart. Brain abscesses are severe neurological infections with high mortality rates. Streptococcus species other than S. pneumoniae are rare causes of brain abscesses. This case report highlights a severe case of extra and intracranial abscesses due to S. constellatus in an immunocompetent host

Meningitis caused by Capnocytophaga canimorsus in a COVID-19 patient: a rare complication of dog bites

Capnocytophaga canimorsus is a gram-negative rod that is part of the commensal microbiota of dogs’ and cats’ mouths. In this case, we report an 85-year-old man with COVID-19 who had his right arm bitten by a dog. His symptoms were impaired consciousness, agitation and aggressive behavior. Physical examination revealed neck stiffness and Brudzinski’s sign. The cerebrospinal fluid culture was compatible with Capnocytophaga canimorsus. He required intensive care and received a 14-day prescription of meropenem. After 40 days of hospitalization, the patient was fully recovered and was discharged. This case highlights the importance of physician and microbiologist be awareness of this disease, mainly in patients with neurological symptoms after a dog or cat bite

Colistin-resistant Escherichia coli belonging to different sequence types: genetic characterization of isolates responsible for colonization, community- and healthcareacquired infections

The plasmid-mediated colistin-resistance gene named mcr-1 has been recently described in different countries and it became a public health challenge. Of note, few studies have addressed the spread of Escherichia coli harboring the mcr-1 gene in both, community and hospital settings. A total of seven colistin-resistant E. coli carrying mcr-1, collected from 2016 to 2018, from community (n=4), healthcare-acquired infections (n=2) and colonization (n=1) were identified in three high complexity hospitals in Sao Paulo, Brazil. These colistin-resistant isolates were screened for mcr genes by PCR and all strains were submitted to Whole Genome Sequencing and the conjugation experiment. The seven strains belonged to seven distinct sequence types (ST744, ST131, ST69, ST48, ST354, ST57, ST10), and they differ regarding the resistance profiles. Transference of mcr-1 by conjugation to E. coli strain C600 was possible in five of the seven isolates. The mcr-1 gene was found in plasmid types IncX4 or IncI2. Three of the isolates have ESBL-encoding genes (blaCTX-M-2, n=2; blaCTX-M-8, n=1). We hereby report genetically distinct E. coli isolates, belonging to seven STs, harboring the mcr-1 gene, associated to community and healthcare-acquired infections, and colonization in patients from three hospitals in Sao Paulo. These findings point out for the potential spread of plasmid-mediated colistin-resistance mechanism in E. coli strains in Brazil

Cotrimoxazole enhances the in vitro susceptibility of Coccidioides posadasii to antifungals

The aim of the present study was to evaluate the effect of cotrimoxazole on the in vitro susceptibility of Coccidioides posadasii strains to antifungals. A total of 18 strains of C. posadasii isolated in Brazil were evaluated in this study. The assays were performed in accordance with the Clinical and Laboratory Standards Institute guidelines and the combinations were tested using the checkerboard method. The minimum inhibitory concentrations were reduced by 11, 2.4, 4.3 and 3.5 times for amphotericin B, itraconazole, fluconazole and voriconazole, respectively. Moreover, it was seen that cotrimoxazole itself inhibited C. posadasii strains in vitro. The impairment of folic acid synthesis may be a potential antifungal target for C. posadasii.Universidade Federal do Ceará Centro Especializado em Micologia MédicaUniversidade Federal do Ceará Programa de Pós-Graduação em Ciências MédicasUniversidade Federal do Ceará Departamento de QuímicaUniversidade Federal do Ceará Departamento de EstatísticaUniversidade Estadual do Ceará Programa de Pós-Graduação em Ciência VeterináriaUniversidade Federal de São Paulo (UNIFESP) Departamento de Microbiologia, Imunologia e ParasitologiaUNIFESP, Depto. de Microbiologia, Imunologia e ParasitologiaSciEL

Aspectos clÃnicos, epidemiolÃgicos e laboratoriais da criptococose no Estado do Cearà entre os anos de 1985 e 2010, bem como efeitos de antirretrovirais inibidores de protease sobre virulÃncia e crescimento de cryptococcus neoformans in vitro

A criptococose à a infecÃÃo causada a partir da inalaÃÃo de leveduras do gÃnero Cryptococcus. A doenÃa costuma acometer mais freqÃentemente pacientes com alguma imunodepressÃo e evoluir de forma subaguda ou crÃnica. Tem predileÃÃo por sistema nervoso central, manifestando-se como meningite, mas tambÃm pode apresentar-se como doenÃa pulmonar ou de forma disseminada, com envolvimento de outros ÃrgÃos. A atividade proteolÃtica em leveduras do gÃnero Cryptococcus tem sido associada a sua patogenicidade. A atividade da enzima fosfolipase tambÃm se faz importante para este gÃnero, uma vez que està relacionada à nutriÃÃo e à capacidade invasiva do microrganismo. Outro mecanismo de virulÃncia de maior relevÃncia para a patogenicidade de Cryptococcus spp. à a cÃpsula polissacarÃdica presente em sua superfÃcie, responsÃvel principal pelo escape aos mecanismos de defesa do hospedeiro. Esta pesquisa teve como objetivo traÃar um perfil clÃnico, epidemiolÃgico e laboratorial dos pacientes com criptococose entre os anos de 1985 e 2010, bem como investigar os efeitos dos inibidores de protease Saquinavir, Darunavir, Ritonavir e Indinavir sobre o crescimento, espessura da cÃpsula, atividade de protease e expressÃo de fosfolipase em cepas de Cryptococcus neoformans isoladas de humanos. A comparaÃÃo entre grupo soropositivo para o HIV com grupo soronegativo mostrou que o primeiro apresenta maiores mÃdias de idade e maior freqÃÃncia de sexo masculino acometido; por outro lado, o grupo soropositivo demonstra menores proteinorraquia e celularidade no lÃquor, bem como menores valores de leucometria e plaquetometria em sangue perifÃrico. ApÃs a realizaÃÃo do teste de sensibilidade por microdiluiÃÃo em caldo, os fungos foram expostos a trÃs concentraÃÃes distintas dos fÃrmacos e avaliados com relaÃÃo à expressÃo das duas enzimas e espessura da cÃpsula. Os resultados mostraram inibiÃÃo significativa (p<0,05) da atividade de protease pelo menos na maior concentraÃÃo testada para todas as drogas, bem como estreitamento da cÃpsula em algumas combinaÃÃes de drogas e cepas. Por outro lado, quanto à atividade de fosfolipase, observou-se um aumento na expressÃo dessa enzima, especialmente nas concentraÃÃes mais elevadas das quatro drogas. Conclui-se, portanto, que apesar de estes antirretrovirais nÃo inibirem o crescimento de Cryptococcus neoformans, sÃo capazes de alterar mecanismos de virulÃncia destas leveduras

Multiresistant Gram-negative microorganisms: mechanisms of resistance and therapeutic alternatives [

Gram negativos multirresistentes têm se tornado uma ameaça em todo o mundo. Os objetivos desse estudo foram apresentar o relato de caso de uma paciente colonizada por microrganismos distintos carreadores do gene mcr-1, descrever sensibilidade de microrganismos multirresistentes a alternativas terapêuticas e seus mecanismos de resistência e avaliar eficácia e segurança do uso de fosfomicina endovenosa. O estudo descreve o caso de uma paciente internada colonizada concomitantemente com E. coli ST744 e K. pneumoniae ST101, ambos portadores de mcr-1, presente em um plasmídeo de 33.304pb. Além disso, 50 microrganismos multirresistentes (14 A. baumannii, 1 P. aeruginosa, 8 S. marcescens e 27 K. pneumoniae), frequentemente produtores de carbapenemase blaKPC-2 (n=28; 56%), blaOXA-23 (n=11; 22%), e outros mecanismos, como enzimas modificadoras de aminoglicosídeos (n=49; 98%), foram avaliados quanto a opções terapêuticas a carbapenêmicos e polimixinas. Destacaram-se os desempenhos de tigeciclina (96% de sensibilidade); minociclina (100%) e doxiciclina (93%) em A. baumannii, ceftazidima/avibactam (96%) em K. pneumoniae e fosfomicina (88%) em S. marcescens. Por último, descrevemos uma série de 13 pacientes com infecções graves tratadas com fosfomicina. Oito pacientes (62%) foram curados. Sinergismo entre fosfomicina e meropenem foi descrito em nove (82%) isolados. Concluiu-se pela possível transmissão in vivo de mcr-1 em espécies distintas, que tigeciclina, minociclina, doxiciclina, ceftazidima/avibactam e fosfomicina podem ser úteis para tratamento de gram negativos multirresistentes, e que fosfomicina é um antimicrobiano seguro e eficaz, especialmente se combinada com o meropenem.Multiresistant gram negatives have become a threat worldwide. The objectives of this study were to present the case report of a patient colonized by distinct microorganisms harboring mcr-1; to describe susceptibility of multiresistant microorganisms to therapeutic alternatives and their mechanisms of resistance; and to evaluate the efficacy and safety of intravenous fosfomycin. The study describes the case of an inpatient colonized concomitantly with E. coli ST744 and K. pneumoniae ST101; both carried mcr-1, present in a 33,304bp plasmid. In addition, 50 multiresistant microorganisms (14 A. baumannii, 1 P. aeruginosa, 8 S. marcescens and 27 K. pneumoniae), that frequently produce carbapenemases blaKPC-2 (n=28; 56%), blaOXA-23 (n= 11; 22%), and other mechanisms such as aminoglycoside modifying enzymes (n=49; 98%), were evaluated for therapeutic alternatives. The performances of tigecycline globally (96% susceptibility); minocycline (100%) and doxycycline (93%) in A. baumannii; as well as ceftazidime/avibactam (96%) in K. pneumoniae, and fosfomycin (88%) in S. marcescens were outstanding. Finally, we described a series of 13 patients with severe infections treated with fosfomycin. Eight patients (62%) were cured. Synergism between fosfomycin and meropenem was described in nine (82%) isolates. We concluded that there is a possible in vivo mcr-1 transmission; that tigecycline, minocycline, doxycycline, ceftazidime/avibactam, and fosfomycin may be helpful for treating gramnegative multiresistant, and that fosfomycin is a safe and effective antimicrobial, especially if combined with meropene

Conjugative transfer of plasmid p_8N_qac(MN687830.1) carrying qacA gene from Staphylococcus aureus to Escherichia coli C600: potential mechanism for spreading chlorhexidine resistance

The methicillin resistant Staphylococcus aureus (MRSA) is recognized by its ability to acquire and transferring resistance genes through interspecies conjugative plasmids. However, transference of plasmids from Gram-positive cocci to Gram-negative bacilli is not well characterized. In this report, we describe the transfer of a conjugative plasmid carrying qacA from MRSA to Escherichia coli C600. We performed a conjugation experiment using a chlorhexidine resistant MRSA isolate (ST-105/SCCmec type III) carrying the gene qacA and qacC as the donor and a chlorhexidine susceptible E. coli C600 isolate as the receptor. Transconjugants were selected using MacConkey agar plates containing chlorhexidine in concentrations ranging from 0.25 to 16 g.L-1. To genotypically confirm the transfer of the resistance gene, the transconjugants were screened by Polymerase Chain Reaction (PCR) and submitted to Sanger’s sequencing. MRSA isolates successfully transferred the chlorhexidine resistance gene (qacA) to the recipient E. coli strain C600. The E. coli transconjugant exhibited an important reduction of chlorhexidine susceptibility, with MICs increasing from ≤ 0.25 to ≥ 16 g.L-1 after conjugation. The qacA gene was detected by PCR as well as in the Sanger’s sequencing analysis of DNA from transconjugant plasmids. To the best of our knowledge, this is the first report of the plasmid p_8N_qac(MN687830.1) carrying qacA and its transfer by conjugation from a MRSA to an E. coli. These findings increase concerns on the emergence of resistance dissemination across the genus and emphasizes the importance of continuous antiseptic stewardship